Bean plant named wills

ABSTRACT

Novel bean plants, such as bean designated WILLS are disclosed. In some embodiments, the disclosure relates to the seeds of bean plant WILLS, to the plants and plant parts of bean WILLS, and to methods for producing a bean plant by crossing the bean plant WILLS with itself or another bean plant. The disclosure further relates to methods for producing other bean plants derived from the bean WILLS.

TECHNICAL FIELD

The present disclosure relates to the field of agriculture, to a new anddistinctive bean plant (Phaseolus vulgaris) designated WILLS and tomethods of making and using such plant.

BACKGROUND

The following description includes information that may be useful inunderstanding the present disclosure. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed disclosures, or that any publication specifically orimplicitly referenced is prior art.

Phaseolus vulgaris, also known as bean, garden bean, or common bean, isan important and valuable vegetable crop. Thus, a continuing goal ofbean plant breeders is to develop stable, high yielding bean plants thatare agronomically sound or unique. The reasons for this goal are tomaximize the amount of yield produced on the land. To accomplish thisgoal, the bean breeder must select and develop bean plants that havetraits that result in superior varieties or cultivars.

SUMMARY

The following embodiments and aspects thereof are described inconjunction with systems, tools and methods which are meant to beexemplary, not limiting in scope.

According to the disclosure, in some embodiments there is provided anovel bean plant designated WILLS, also interchangeably referred to as‘bean plant WILLS,’ ‘bean WILLS,’ ‘common bean plant WILLS,’ or ‘gardenbean plant WILLS.’

This disclosure thus relates to the seeds of bean plant designatedWILLS, to the plants or parts of bean plant designated WILLS, to plantsor parts of bean plant comprising all of the physiological andmorphological characteristics of bean plant designated WILLS and/orhaving all of the physiological and morphological characteristics ofbean plant designated WILLS, and/or having one or more of or all of thecharacteristics of bean plant designated WILLS including but not limitedto as determined at the 5% significance level when grown in the sameenvironmental conditions, and/or having one or more of the physiologicaland morphological characteristics of bean plant designated WILLSincluding but not limited to as determined at the 5% significance levelwhen grown in the same environmental conditions and/or having all of thephysiological and morphological characteristics of bean plant designatedWILLS including but not limited to as determined at the 5% significancelevel when grown in the same environmental conditions and/or having oneor more of the physiological and morphological characteristics of beanplant designated WILLS when grown in the same environmental conditionsand/or having all of the physiological and morphological characteristicsof bean plant designated WILLS when grown in the same environmentalconditions. The disclosure also relates to variants, mutants and trivialmodifications of bean plant designated WILLS and parts thereof. In someembodiments, a representative sample of seed of bean plant designatedWILLS is deposited under NCIMB No. ______.

Plant parts of the bean plant designated WILLS of the present disclosureare also provided, such as a pod and a bean, but not limited to, a seed,a bean, a scion, a rootstock, a pod, a leaf, a flower, a peduncle, astalk, a root, a stamen, an anther, a pistil, a pollen or an ovuleobtained from the cultivar. The present disclosure provides pods of thebean plant designated WILLS of the present disclosure. Such pods and/orbeans could be used as fresh products for consumption or in processesresulting in processed products such as food products comprising one ormore harvested part of the bean plant WILLS, such as prepared podsand/or beans or parts thereof, canned pods and/or beans or partsthereof, freeze-dried or frozen pods and/or beans or parts thereof,diced pods and/or beans, squeezed pods and/or beans, juices of podsand/or beans, prepared cuts of pods and/or beans, canned pods and/orbeans. All such products are part of the present disclosure and thelike. The harvested parts or food products can be or can comprise thebean pods of the bean plant WILLS. The food products might haveundergone one or more processing steps such as, but not limited tocutting, washing, mixing, frizzing, canning, etc. All such products arepart of the present disclosure. The present disclosure also providesplant parts or cells of the bean plant designated WILLS, wherein a plantregenerated from said plants parts or cells has one or more of, or allthe phenotypic and morphological characteristics of bean plantdesignated WILLS, such as one or more of or all the characteristics ofbean plant designated WILLS deposited under NCIMB No. ______. All suchparts and cells are part of the present disclosure.

The plants and seeds of the present disclosure include those that may beof an essentially derived variety as defined in section 41(3) of thePlant Variety Protection Act of The United States of America, e.g., avariety that is predominantly derived from bean plant designated WILLSor from a variety that i) is predominantly derived from bean plantdesignated WILLS, while retaining the expression of the essentialcharacteristics that result from the genotype or combination ofgenotypes of bean plant designated WILLS; ii) is clearly distinguishablefrom bean plant designated WILLS; and iii) except for differences thatresult from the act of derivation, conforms to the initial variety inthe expression of the essential characteristics that result from thegenotype or combination of genotypes of the bean plant designated WILLS.

In another aspect, the present disclosure provides regenerable cells. Insome embodiments, the regenerable cells are for use in tissue culture ofbean plant designated WILLS. In some embodiments, the tissue culture iscapable of regenerating plants comprising all of the physiological andmorphological characteristics of bean plant designated WILLS, and/orhaving all of the physiological and morphological characteristics ofbean plant designated WILLS, and/or having one or more of thephysiological and morphological characteristics of bean plant designatedWILLS, and/or having the characteristics of bean plant designated WILLS.In some embodiments, the regenerated plants have the characteristics ofbean plant designated WILLS including but not limited to as determinedat the 5% significance level when grown in the same environmentalconditions and/or have all of the physiological and morphologicalcharacteristics of bean plant designated WILLS including but not limitedto as determined at the 5% significance level when grown in the sameenvironmental conditions and/or have one or more of the physiologicaland morphological characteristics bean plant designated WILLS includingbut not limited to as determined at the 5% significance level when grownin the same environmental conditions and/or having all of thephysiological and morphological characteristics of bean plant designatedWILLS when grown in the same environmental conditions.

In some embodiments, the plant parts and cells used to produce suchtissue cultures will be embryos, meristematic cells, seeds, callus,pollens, leaves, anthers, pistils, stamens, roots, root tips, stems,petioles, pods, cotyledons, hypocotyls, ovaries, seed coats, pods,stalks, endosperms, flowers, axillary buds or the like. Protoplastsproduced from such tissue culture are also included in the presentdisclosure. The bean leaves, shoots, roots and whole plants regeneratedfrom the tissue culture, as well as the pods and/or beans produced bysaid regenerated plants are also part of the disclosure. In someembodiments, the whole plants regenerated from the tissue culture haveone, more than one, or all of the physiological and morphologicalcharacteristics of bean plant designated WILLS, including but notlimited to as determined at the 5% significance level when grown in thesame environmental conditions.

The disclosure also provides for methods for vegetatively propagating aplant of the present disclosure. In the present application,vegetatively propagating can be interchangeably used with vegetativereproduction. In some embodiments, the methods comprise collecting partsof a bean plant designated WILLS and regenerating a plant from saidparts. In some embodiments, one of the parts can be, for example, astem. In some embodiments, the methods can be, for example, a stemcutting that is rooted into an appropriate medium according totechniques known by the one skilled in the art. Plants and partsthereof, including but not limited to pods thereof, produced by suchmethods are also included in the present disclosure. In another aspect,the plants and pods thereof such as stems and pods produced by suchmethods comprise all of the physiological and morphologicalcharacteristics of bean plant designated WILLS, and/or have all of thephysiological and morphological characteristics of bean plant designatedWILLS and/or have the physiological and morphological characteristics ofbean plant designated WILLS and/or have one or more of thecharacteristics of bean plant designated WILLS. In some embodiments,plants, parts or pods/beans thereof produced by such methods consist ofone, more than one, or all of the physiological and morphologicalcharacteristics of bean plant designated WILLS, including but notlimited to as determined at the 5% significance level when grown in thesame environmental conditions.

Further included in the disclosure are methods for producing pods,including beans, from the bean plant designated WILLS. In someembodiments, the methods comprise growing a bean plant designated WILLSto produce a bean pod and/or beans. In some embodiments, the methodsfurther comprise harvesting the bean pod. Such bean pods, beans, and/orbeans thereof are parts of the present disclosure. In some embodiments,such pods, beans, and/or seeds have all of the physiological andmorphological characteristics of the pods, beans, and/or seeds of beanplant designated WILLS (e.g. those listed in Table 1 and/or depositedunder NCIMB No. ______) when grown in the same environmental conditionsand/or have one or more of the physiological and morphologicalcharacteristics of the pods, beans, and/or seeds of the bean plantdesignated WILLS (e.g. those listed in Table 1 and/or deposited underNCIMB No. ______) when grown in the same environmental conditions and/orhave the characteristics of the pods, beans, and/or seeds of the beanplant designated WILLS (e.g. those listed in Table 1 and/or depositedunder NCIMB No. ______) when grown in the same environmental conditions.

Also included in this disclosure are methods for producing a bean plant.In some embodiments, the bean plant is produced by crossing the beanplant designated WILLS with itself or another bean plant. In someembodiment, the other plant can be a bean variety/cultivar/line otherthan WILLS, a bean hybrid or a plant of Phaseolus genus. When crossedwith itself, i.e., when WILLS is crossed with another bean plant WILLSor self-pollinated, bean plant WILLS will be conserved (e.g. as aninbred). When crossed with another, different bean plant, an F1 hybridseed is produced if the different bean plant is an inbred and a“three-way cross” seed is produced if the different bean plant is ahybrid. Such F1 hybrid seed and three-way hybrid seeds and plantsproduced by growing said F1 and three-way hybrid seeds are included inthe present disclosure. Methods for producing a F1 and three-way hybridbean seed comprising crossing bean plant WILLS with a different beanvariety/cultivar/line or hybrid and harvesting the resultant hybrid beanseed are also part of the disclosure. The bean seeds produced by themethods comprising crossing bean plant WILLS bean plant with a differentbean plant and harvesting the resultant bean seed are included in thedisclosure, as are included the bean plant or parts thereof and seedsproduced by said grown bean plants.

Further included in the disclosure are methods for producing a beanseeds and plants made thereof. In some embodiments, the methods compriseself-pollinating the bean plant designated WILLS and harvesting theresultant seeds. Bean seeds produced by such method are also part of thedisclosure.

In another embodiment, this disclosure relates to methods for producingother bean plant designated WILLS from a collection of seeds.

In some embodiments, the collection contains both seeds of inbred parentline(s) of bean plant designated WILLS seeds. Such a collection of seedsmight be a commercial bag of seeds. In some embodiments, said methodscomprise planting the collection of seeds. When planted, the collectionof seeds will produce inbred parent lines of bean plant WILLS and plantsfrom the seeds of WILLS. In some embodiments, said inbred parent linesof bean plant designated WILLS plants are identified as having adecreased vigor compared to the other plants (i.e., plants) grown fromthe collection of seeds. In some embodiments, said decreased vigor isdue to the inbreeding depression effect and can be identified forexample by a less vigorous appearance for vegetative and/or reproductivecharacteristics including a shorter plant height, small pod size, podshape, pod color or other characteristics. In some embodiments, seeds ofthe inbred parent lines of the bean plant WILLS are collected and, ifnew inbred parent plants thereof are grown and crossed in a controlledmanner with each other, the bean plant WILLS will be recreated.

This disclosure also relates to methods for producing other bean plantsderived from bean plant WILLS and to the bean plants derived by the useof methods described herein.

In some embodiments, such methods for producing a bean plant derivedfrom bean plant WILLS comprise (a) self-pollinating the bean plant WILLSat least once to produce a progeny plant derived from the bean plantWILLS. In some embodiments, the methods further comprise (b) crossingthe progeny plant derived from the bean plant WILLS with itself or asecond bean plant to produce a seed of a progeny plant of a subsequentgeneration. In some embodiments, the methods further comprise (c)growing the progeny plant of the subsequent generation. In someembodiments, the methods further comprise (d) crossing the progeny plantof the subsequent generation with itself or a second g bean plant toproduce a bean plant further derived from the bean plant WILLS. Infurther embodiments, steps (b), step (c) and/or step (d) are repeatedfor at least 1, 2, 3, 4, 5, 6, 7, 8, or more generations to produce abean plant derived from the bean plant WILLS. In some embodiments,within each crossing cycle, the second plant is the same plant as thesecond plant in the last crossing cycle. In some embodiments, withineach crossing cycle, the second plant is different from the second plantin the last crossing cycle, which can be a bean variety/cultivar/lineother than WILLS, a bean hybrid or a plant of Phaseolus genus.

Another method for producing a bean plant derived from bean plant WILLS,comprises (a) crossing the bean plant WILLS plant with a second beanplant to produce a progeny plant derived from the bean plant WILLS. Insome embodiments, the method further comprises (b) crossing the progenyplant derived from the bean plant WILLS with itself or a second beanplant to produce a seed of a progeny plant of a subsequent generation.In some embodiments, the method further comprises (c) growing theprogeny plant of the subsequent generation. In some embodiments, themethod further comprises (d) crossing the progeny plant of thesubsequent generation with itself or a second bean plant to produce abean plant derived from the bean plant WILLS. In a further embodiment,steps (b), (c) and/or (d) are repeated for at least 1, 2, 3, 4, 5, 6, 7,8, or more generations to produce a bean plant derived from the beanplant WILLS. In some embodiments, within each crossing cycle, the secondplant is the same plant as the second plant in the last crossing cycle.In some embodiments, within each crossing cycle, the second plant isdifferent from the second plant in the last crossing cycle, which can bea bean variety/cultivar/line other than WILLS, a bean hybrid or a plantof Phaseolus genus.

In one aspect, the present disclosure provides methods of introducing asingle locus conversion conferring one or more desired trait(s) into thebean plant WILLS, and plants or parts including pods and/or seedsobtained from such methods. In another aspect, the present disclosureprovides methods of modifying a single locus and conferring one or moredesired trait(s) into the bean plant WILLS, and plants or partsincluding pods and/or seeds, and/or bean, obtained from such methods.The desired trait(s) may be, but not exclusively, conferred by a singlelocus that contains a single and/or multiple gene(s). In someembodiments, the gene is a dominant allele. In some embodiments, thegene is a partially dominant allele. In some embodiments, the gene is arecessive allele. In some embodiments, the gene or genes will confer ormodify such traits, including but not limited to male sterility,herbicide resistance, insect resistance, resistance for bacterial,fungal, mycoplasma or viral disease, enhanced plant quality such asimproved drought or salt tolerance, water-stress tolerance, improvedstandability, enhanced plant vigor, improved shelf life, delayedsenescence or controlled ripening, enhanced nutritional quality such asincreased sugar content or increased sweetness, increased texture,flavor and aroma, improved pod length and/or size, protection for color,pod shape, uniformity, length or diameter, refinement or depth, lodgingresistance, yield and recovery, improve fresh cut application, specificaromatic compounds, specific volatiles, flesh texture and specificnutritional components. For the present disclosure and the skilledartisan, disease is understood to include, but not limited to fungaldiseases, viral diseases, bacterial diseases, mycoplasma diseases, orother plant pathogenic diseases and a disease resistant plant willencompass a plant resistant to fungal, viral, bacterial, mycoplasma, andother plant pathogens. In one aspect, the gene or genes may be naturallyoccurring bean gene(s) and/or spontaneous or induced mutations(s). Inanother aspect, genes are mutated, modified, genetically engineeredthrough the use of New Breeding Techniques described herein. In someembodiments, the method for introducing the desired trait(s) into beanplant WILLS is a backcrossing process by making use of a series ofbackcrosses to bean plant WILLS or at least one of the parental inbredlines of bean plant designated WILLS, which the desired trait(s) ismaintained by selection. Bean plant WILLS or at least one of theparental inbred lines of bean plant designated WILLS possesses thedesired trait(s) by the backcrossing process, and the desired trait(s)is inherited by the bean progeny plants by conventional breedingtechniques known to breeders of ordinary skill in the art. The singlegene converted plants or single locus converted plants that can beobtained by the methods are included in the present disclosure.

When dealing with a gene that has been modified, for example through NewBreeding Techniques, the trait (genetic modification) could be directlymodified into the newly developed bean plant and/or at least one of theparental inbred lines of bean plant WILLS. Alternatively, if the traitis not modified into each newly developed bean plant and/or at least oneof the parent lines of bean plant WILLS, another typical method used bybreeders of ordinary skill in the art to incorporate the modified geneis to take a line already carrying the modified gene and to use suchline as a donor line to transfer the modified gene into the newlydeveloped bean plant and/or at least one of the parent lines of thenewly developed plant. The same would apply for a naturally occurringtrait or one arising from spontaneous or induced mutations.

In some embodiments, the backcross breeding process of bean plant WILLScomprises (a) crossing one of the parental inbred line plants of beanplant WILLS with plants of another line that comprise the desiredtrait(s) to produce F₁ progeny plants. In some embodiments, the processfurther comprises (b) selecting the F₁ progeny plants that have thedesired trait(s). In some embodiments, the process further comprises (c)crossing the selected F₁ progeny plants with the parental inbred beanlines of bean WILLS plants to produce backcross progeny plants. In someembodiments, the process further comprises (d) selecting for backcrossprogeny plants that have the desired trait(s) and essentially all of thephysiological and morphological characteristics of the bean parentalinbred line of bean plant WILLS to produce selected backcross progenyplants. In some embodiments, the process further comprises (e) repeatingsteps (c) and (d) one, two, three, four, five six, seven, eight, nine ormore times in succession to produce selected, second, third, fourth,fifth, sixth, seventh, eighth, ninth or higher backcross progeny plantsthat have the desired trait(s) and essentially all of thecharacteristics of the bean plant WILLS, and/or have the desiredtrait(s) and essentially all of the physiological and morphologicalcharacteristics of the parental inbred line of bean plant WILLS, and/orhave the desired trait(s) and otherwise essentially all of thephysiological and morphological characteristics of the bean plant WILLS,including but not limited to when grown in the same environmentalconditions or including but not limited to at a 5% significance levelwhen grown in the same environmental conditions. The bean plants or seedproduced by the methods are also part of the disclosure, as are the beanWILLS plants that comprised the desired trait. Backcrossing breedingmethods, well known to one skilled in the art of plant breeding will befurther developed in subsequent parts of the specification.

An embodiment of this disclosure is a method of making a backcrossconversion of bean plant WILLS. In some embodiments, the methodcomprises crossing bean plant WILLS with a donor plant comprising aninduced gene mutation(s), a naturally occurring gene mutation(s) or agene(s) and/or sequence(s) modified through New Breeding Techniquesconferring one or more desired traits to produce F₁ progeny plants. Insome embodiments, the method further comprises selecting an F₁ progenyplant comprising the naturally occurring gene mutation(s), induced genemutation(s) or gene(s) and/or sequences(s) modified through New BreedingTechniques conferring the one or more desired traits. In someembodiments, the method further comprises backcrossing the selectedprogeny plant to the bean plant WILLS. This method may further comprisethe step of obtaining a molecular marker profile of the bean plant WILLSand using the molecular marker profile to select for the progeny plantwith the desired trait and the molecular marker profile of the beanplant WILLS. In some embodiments, this method further comprises crossingthe backcross progeny plant of the parental bean plant WILLS containingthe naturally occurring gene mutation(s), the induced gene mutation(s)or the gene(s) and or sequences modified through New Breeding Techniquesconferring the one or more desired traits with the second parental beanplants of bean plant WILLS in order to produce the bean plant WILLScomprising the naturally occurring gene mutation(s), the induced genemutation(s) or the gene(s) and/or sequences modified through NewBreeding Techniques conferring the one or more desired traits. Theplants or parts thereof produced by such methods are also part of thepresent disclosure.

In some embodiments of the disclosure, the number of loci that may betransferred and/or backcrossed into the bean plant WILLS is at least 1,2, 3, 4, 5, or more.

A single locus may contain several genes. A single locus conversion alsoallows for making one or more site specific changes to the plant genome,such as, without limitation, one or more nucleotide changes, deletions,insertions, substitutions, etc. In some embodiments, the single locusconversion is performed by genome editing, a.k.a. genome editing withengineered nucleases (GEEN). In some embodiments, the genome editingcomprises using one or more engineered nucleases. In some embodiments,the engineered nucleases include, but are not limited to Zinc fingernucleases (ZFNs), Transcription Activator-Like Effector Nucleases(TALENs), the CRISPR/Cas system (using such as Cas9, Cas12a/Cpf1,Cas13/C2c2, CasX and CasY), engineered meganuclease, engineered homingendonucleases and endonucleases for DNA guided genome editing (Gao etal., Nature Biotechnology (2016), doi: 10.1038/nbt.3547). In someembodiments, the single locus conversion changes one or severalnucleotides of the plant genome. Such genome editing techniques are someof the techniques now known by the person skilled in the art and hereinare collectively referred to as “New Breeding Techniques”. In someembodiments, one or more above-mentioned genome editing methods aredirectly applied on a plant of the present disclosure, rather than onthe parental bean lines of WILLS. Accordingly, a cell containing anedited genome, or a plant part containing such cell can be isolated andused to regenerate a novel plant which has a new trait conferred by saidgenome editing, and essentially all of the physiological andmorphological characteristics of bean plant WILLS.

The disclosure further provides methods for developing bean plants in abean plant breeding program using plant breeding techniques includingbut not limited to, recurrent selection, backcrossing, pedigreebreeding, genomic selection, molecular marker (Isozyme Electrophoresis,Restriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reactions(AP-PCRs), DNA Amplification Fingerprintings (DAFs), SequenceCharacterized Amplified Regions (SCARs), Amplified Fragment LengthPolymorphisms (AFLPs), and Simple Sequence Repeats (SSRs) which are alsoreferred to as Microsatellites, Single Nucleotide Polymorphisms (SNPs),enhanced selection, genetic markers, enhanced selection andtransformation. Seeds, pods, bean plants, and parts thereof produced bysuch breeding methods are also part of the disclosure.

The disclosure also relates to variants, mutants and trivialmodifications of the bean plant WILLS, parts thereof or parental inbredlines thereof. Variants, mutants and trivial modifications of bean plantWILLS and parts (such as seeds, pods, beans etc.) thereof can begenerated by methods available to one skilled in the art, including butnot limited to, mutagenesis (e.g., chemical mutagenesis, radiationmutagenesis, transposon mutagenesis, insertional mutagenesis, signaturetagged mutagenesis, site-directed mutagenesis, and natural mutagenesis),knock-outs/knock-ins, antisense oligonucleotides, RNA interference andother techniques such as the New Breeding Techniques described herein.For more information of mutagenesis in plants, such as agents orprotocols, see Acquaah et al. (Principles of plant genetics andbreeding, Wiley-Blackwell, 2007, ISBN 1405136464, 9781405136464, whichis herein incorporated by reference in its entity).

The disclosure also relates to a mutagenized population of the beanplant WILLS and methods of using such populations. In some embodiments,the mutagenized population can be used in screening for new bean plantswhich comprise essentially one or more of or all the morphological andphysiological characteristics of bean plant WILLS. In some embodiments,the new bean plants obtained from the screening process compriseessentially all of the morphological and physiological characteristicsof the bean plant WILLS, and one or more additional or differentmorphological and physiological characteristics that the bean plantWILLS does not have.

This disclosure is also directed to methods for producing a bean plantby crossing a first parent bean plant with a second parent bean plant,wherein either the first or second parent bean plant is a bean plant ofWILLS. Further, both first and second parent bean plants can come fromthe bean plant WILLS. Further, the bean plant WILLS can beself-pollinated i.e., the pollen of a bean plant WILLS can pollinate theovule of the same bean plant WILLS. When crossed with another beanplant, a seed is produced. Such methods of hybridization andself-pollination are well known to those skilled in the art of breeding.

A bean plant such as one of the parental lines of bean plant WILLS hasbeen produced through several cycles of self-pollination and istherefore to be considered as a homozygous line. An inbred line can alsobe produced though the dihaploid system which involves doubling thechromosomes from a haploid plant or embryo thus resulting in an inbredline that is genetically stable (homozygous) and can be reproducedwithout altering the inbred line. Haploid plants could be obtained fromhaploid embryos that might be produced from microspores, pollen, anthercultures or ovary cultures or spontaneous haploidy. The haploid embryosmay then be doubled by chemical treatments such as by colchicine or bedoubled autonomously. The haploid embryos may also be grown into haploidplants and treated to induce the chromosome doubling. In either case,fertile homozygous plants may be obtained. A variety is classicallycreated through the fertilization of an ovule from an inbred parentalline by the pollen of another, different inbred parental line. Due tothe homozygous state of the inbred line, the produced gametes carry acopy of each parental chromosome. As both the ovule and the pollen bringa copy of the arrangement and organization of the genes present in theparental lines, the genome of each parental line is present in theresulting F₁, theoretically in the arrangement and organization createdby the plant breeder in the original parental line.

As long as the homozygosity of the parental lines is maintained, theresulting cross shall be stable. The F₁ is then a combination ofphenotypic characteristics issued from two arrangement and organizationof genes, both created by a person skilled in the art through thebreeding process.

Still further, this disclosure is also directed to methods for producinga bean plant WILLS-derived bean plant by crossing bean plant WILLS witha second bean plant. In some embodiments, the methods further compriseobtaining a progeny seed from the cross. In some embodiments, themethods further comprise growing the progeny seed, and possiblyrepeating the crossing and growing steps with the bean plant WILLSderived plant from 0 to 7 or more times. Thus, any such methods usingthe bean plant WILLS are part of this disclosure: selfing, backcrosses,production, crosses to populations, and the like. All plants producedusing bean plant WILLS as a parent are within the scope of thisdisclosure, including plants derived from bean plant WILLS. In someembodiments, such plants have one, more than one or all of thephysiological and morphological characteristics of the bean plant WILLSincluding but not limited to as determined at the 5% significance levelwhen grown in the same environmental conditions. In some embodiments,such plants might exhibit additional and desired characteristics ortraits such as high seed yield, high seed germination, seedling vigor,early maturity, high pod yield, ease of pod setting, disease toleranceor resistance, lodging resistance, and adaptability for soil and climateconditions. Consumer-driven traits, such as a preference for a given podsize, pod shape, pod color, pod texture, pod taste, pod firmness, podsugar content are other traits that may be incorporated into new beanplants developed by this disclosure.

A bean plant can also be propagated vegetatively. A part of the plant,for example a shoot tissue, is collected, and a new plant is obtainedfrom the part. Such part typically comprises an apical meristem of theplant. The collected part is transferred to a medium allowingdevelopment of a plantlet, including for example rooting or developmentof shoots, or is grafted onto a bean plant or a rootstock prepared tosupport growth of shoot tissue. This is achieved using methods wellknown in the art. Accordingly, in one embodiment, a method ofvegetatively propagating a plant of the present disclosure comprisescollecting a part of a plant according to the present disclosure, e.g. ashoot tissue, and obtaining a plantlet from said part. In oneembodiment, a method of vegetatively propagating a plant of the presentdisclosure comprises: (a) collecting tissue of a plant of the presentdisclosure; (b) rooting said proliferated shoots to obtain rootedplantlets. In one embodiment, a method of vegetatively propagating aplant of the present disclosure comprises: (a) collecting tissue of aplant of the present disclosure; (b) cultivating said tissue to obtainproliferated shoots; (c) rooting said proliferated shoots to obtainrooted plantlets. In one embodiment, such method further comprisesgrowing a plant from said plantlets. In one embodiment, a pod isharvested from said plant. In one embodiment, such pods and plants haveall of the physiological and morphological characteristics of pods andplants of bean plant designated WILLS when grown in the sameenvironmental conditions. In one embodiment, the pod is processed intoproducts such as canned bean pods and/or parts thereof, freeze dried orfrozen pod and/or parts thereof, fresh or prepared pod and/or partsthereof or pastes, sauces, purees and the like.

The disclosure is also directed to the use of the bean plant WILLS in agrafting process. In one embodiment, the bean plant WILLS is used as thescion while in another embodiment, the bean plant WILLS is used as arootstock.

In some embodiments, the present disclosure teaches a seed of bean plantdesignated WILLS, wherein a representative sample of seed of said beanplant is deposited under NCIMB No. ______.

In some embodiments, the present disclosure teaches a bean plant, or apart thereof, produced by growing the deposited WILLS seed.

In some embodiments, the present disclosure teaches a bean plant part,wherein the bean part is selected from the group consisting of: a leaf,a flower, a pod, a bean, a stalk, a root, a rootstock, a seed, anembryo, a peduncle, a stamen, an anther, a pistil, an ovule, a pollen, acell, and a scion.

In some embodiments, the present disclosure teaches a bean plant, or apart thereof, having all of the characteristics of bean plant WILLSdeposited under NCIMB No. ______ of this disclosure including but notlimited to as determined at the 5% significance level when grown in thesame environmental conditions.

In some embodiments, the present disclosure teaches a bean plant, or apart thereof, having all of the physiological and morphologicalcharacteristics of bean plant WILLS, wherein a representative sample ofseed of said bean plant was deposited under NCIMB No. ______.

In some embodiments, the present disclosure teaches a tissue culture ofregenerable cells produced from the plant or part grown from thedeposited bean plant WILLS seed, wherein cells of the tissue culture areproduced from a plant part selected from the group consisting ofprotoplasts, embryos, meristematic cells, callus, pods, beans, pollens,ovules, flowers, seeds, leaves, roots, root tips, anthers, stems,petioles, pods, axillary buds, cotyledons and hypocotyls. In someembodiments, the plant part includes protoplasts produced from a plantgrown from the deposited bean plant WILLS seed.

In some embodiments, the present disclosure teaches a compositioncomprising regenerable cells produced from the plant or part thereofgrown from the deposited WILLS seed, or other part or cell thereof. Insome embodiments, the composition further comprises a growth media. Insome embodiments, the growth media is solid or a synthetic cultivationmedium. In some embodiments, the composition is a bean plant regeneratedfrom the tissue culture from a plant grown from the deposited bean plantWILLS seed, said plant having all of the characteristics of bean plantWILLS, wherein a representative sample of seed of said bean plant isdeposited under NCIMB No. ______.

In some embodiments, the present disclosure teaches a bean pod and/orbeans produced from the plant grown from the deposited bean plant WILLSseed.

In some embodiments, such pods have all of the physiological andmorphological characteristics of pods of bean plant designated WILLSwhen grown in the same environmental conditions.

In some embodiments, methods of producing said bean pod and/or seedcomprise a) growing the bean plant from deposited bean plant WILLS seedto produce a bean pod, and (b) harvesting said bean pod and/or seed. Insome embodiments, the present disclosure also teaches a bean pod and/orseed produced by the method of producing bean pod and/or seed asdescribed above. In some embodiments, such pods have all of thephysiological and morphological characteristics of pods of bean plantdesignated WILLS (e.g. those listed in Table 1 and/or deposited underNCIMB No. ______) when grown in the same environmental conditions.

In some embodiments, the present disclosure teaches methods forproducing a bean seed comprising crossing a first parent bean plant witha second parent bean plant and harvesting the resultant bean seed,wherein said first parent bean plant and/or second parent bean plant isthe bean plant produced from the deposited bean plant WILLS seed or abean plant having all of the characteristics of bean plant WILLSdeposited under NCIMB No. ______ including but not limited to asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

In some embodiments, the present disclosure teaches methods forproducing a bean seed comprising self-pollinating the bean plant grownfrom the deposited bean plant WILLS seed and harvesting the resultantbean seed.

In some embodiments, the present disclosure teaches the seed produced byany of the above described methods.

In some embodiments, the present disclosure teaches methods ofvegetatively propagating the bean plant grown from the deposited beanplant WILLS seed, said method comprising a) collecting a part of a plantgrown from the deposited bean WILLS seed and regenerating a plant fromsaid part.

In some embodiments, the method further comprises harvesting pods and/orseeds from said vegetatively propagated plant. In some embodiments, themethod further comprises harvesting a pod and/or a seed from saidvegetatively propagated plant.

In some embodiments, the present disclosure teaches the plant, parts,pods and/or seeds of plants vegetatively propagated from parts of plantsgrown from the deposited bean WILLS seed. In some embodiments, suchplant, parts, pods and/or seeds have all of the physiological andmorphological characteristics of plant, parts, pods and/or seeds of beanplant WILLS (e.g. those listed in Table 1 and/or deposited under NCIMBNo. ______) when grown in the same environmental conditions.

In some embodiments, the present disclosure teaches methods of producinga bean plant derived from the bean plant WILLS. In some embodiment, themethods comprise (a) self-pollinating the plant grown from the depositedbean plant WILLS seed at least once to produce a progeny plant derivedfrom bean plant WILLS. In some embodiments, the method further comprises(b) crossing the progeny plant derived from bean plant WILLS with itselfor a second bean plant to produce a seed of a progeny plant of asubsequent generation; and; (c) growing the progeny plant of thesubsequent generation from the seed, and (d) crossing the progeny plantof the subsequent generation with itself or a second bean plant toproduce a bean plant derived from the bean plant WILLS. In someembodiments said methods further comprise the step of: (e) repeatingsteps (b), (c) and/or (d) for at least 1, 2, 3, 4, 5, 6, 7, or moregeneration to produce a bean plant derived from the bean plant WILLS.

In some embodiments, the present disclosure teaches methods of producinga bean plant derived from the bean plant WILLS, the methods comprising(a) crossing the plant grown from the deposited bean plant WILLS seedwith a second garden bean plant to produce a progeny plant derived frombean plant WILLS. In some embodiments, the method further comprises; (b)crossing the progeny plant derived from bean WILLS with itself or asecond bean plant to produce a seed of a progeny plant of a subsequentgeneration; and; (c) growing the progeny plant of the subsequentgeneration from the seed; (d) crossing the progeny plant of thesubsequent generation with itself or a second bean plant to produce abean plant derived from the bean plant WILLS. In some embodiments saidmethods further comprise the steps of: (e) repeating steps (b), (c)and/or (d) for at least 1, 2, 3, 4, 5, 6, 7 or more generations toproduce a bean plant derived from the bean plant WILLS.

In some embodiments, the present disclosure teaches plants grown fromthe deposited bean plant WILLS seed wherein said plants comprise asingle locus conversion. As used herein, the term “a” or “an” refers toone or more of that entity; for example, “a single locus conversion”refers to one or more single locus conversions or at least one singlelocus conversion. As such, the terms “a” (or “an”), “one or more” and“at least one” are used interchangeably herein. In addition, referenceto “an element” by the indefinite article “a” or “an” does not excludethe possibility that more than one of the elements are present, unlessthe context clearly requires that there is one and only one of theelements.

In some embodiments, the present disclosure teaches a method ofproducing a plant of bean plant designated WILLS comprising at least onedesired trait, the method comprising introducing a single locusconversion conferring the desired trait into bean plant designatedWILLS, whereby a plant of bean designated WILLS comprising the desiredtrait is produced.

In some embodiments, the present disclosure teaches a bean plant,comprising a single locus conversion and essentially all of thecharacteristics of bean plant designated WILLS when grown under the sameenvironmental conditions, wherein a representative sample of seed ofsaid bean plant has been deposited under NCIMB No. ______. In otherembodiments, the single locus conversion is introduced into the plant bythe use of recurrent selection, mutation breeding, wherein said mutationbreeding selects for a mutation that is spontaneous or artificiallyinduced, backcrossing, pedigree breeding, haploid/double haploidproduction, marker-assisted selection, genetic transformation, genomicselection, Zinc finger nuclease (ZFN) technology, oligonucleotidedirected mutagenesis, cisgenesis, intragenesis, RNA-dependent DNAmethylation, agro-infiltration, Transcription Activation-Like EffectorNuclease (TALENs), CRISPR/Cas system, engineered meganuclease,re-engineered homing endonuclease, and DNA guided genome editing.

In some embodiments, the plant comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,or more single locus conversions. In some embodiments, the plantcomprises no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 single locusconversions, but essentially all of the other physiological andmorphological characteristics of bean plant WILLS and/or deposited underNCIMB No. ______. In some embodiments, the plant comprises at least onesingle locus conversion and essentially all of the physiological andmorphological characteristics of bean plant WILLS deposited under NCIMBNo. ______. In other embodiments, the plant comprises one single locusconversion and essentially all of the other physiological andmorphological characteristics of bean plant WILLS deposited under NCIMBNo. ______.

In some embodiments, said single locus conversion confers said plantswith a trait selected from the group consisting of male sterility, malefertility, herbicide resistance, insect resistance, resistance forbacterial, fungal, mycoplasma or viral disease, enhanced plant qualitysuch as improved drought or salt tolerance, water stress tolerance,improved standability, enhanced plant vigor, improved shelf life,delayed senescence or controlled ripening, increased nutritional qualitysuch as increased sugar content or increased sweetness, increasedtexture, flavor and aroma, improved pod length and/or size, protectionfor color, pod shape, uniformity, length or diameter, refinement ordepth lodging resistance, yield and recovery when compared to a suitablecheck/comparison plant. In further embodiments, the single locusconversion confers said plant with herbicide resistance.

In some embodiments, the check plant is a bean plant not having saidsingle locus conversion conferring the desired trait(s). In someembodiments, the at least one single locus conversion is anaturally-occurring mutation, an artificially mutated gene, or a gene ornucleotide sequence modified through the use of New Breeding Techniques.

In some embodiments, the present disclosure teaches methods of producinga bean plant, comprising grafting a rootstock or a scion of the beanplant grown from the deposited WILLS seed to another bean plant. In someembodiments, the present disclosure teaches methods for producingnucleic acids, comprising isolating nucleic acids from the plant grownfrom the deposited WILLS seed, or a part, or a cell thereof. In someembodiments, the present disclosure teaches methods for producing asecond bean plant, comprising applying plant breeding techniques to theplant grown from the deposited WILLS seed, or part thereof to producethe second bean plant.

In some embodiments, the present disclosure provides a method ofproducing a commodity plant product comprising collecting the commodityplant product from the plant of the present disclosure. The commodityplant product produced by said method is also part of the presentdisclosure.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DETAILED DESCRIPTION Definitions

In the description and tables that follow, a number of terms are used.In order to provide a clear and consistent understanding of thespecification and claims, including the scope to be given such terms,the following definitions are provided:

Allele: An allele is any of one or more alternative forms of a genewhich relate to one trait or characteristic. In a diploid cell ororganism, the two alleles of a given gene occupy corresponding loci on apair of homologous chromosomes.

Backcrossing: Backcrossing is a process in which a breeder repeatedlycrosses hybrid progeny back to one of the parents, for example, a firstgeneration hybrid F₁ with one of the parental genotype of the F₁ hybrid.

Bean yield (tons/acre): The yield in tons/acre is the actual yield ofthe bean pods at harvest.

Commodity plant product: A “commodity plant product” refers to anycomposition or product that is comprised of material derived from aplant, seed, plant cell, or plant part of the present disclosure.Commodity plant products may be sold to consumers and can be viable ornonviable. Nonviable commodity products include but are not limited tononviable seeds and grains; processed seeds, seed parts, and plantparts; dehydrated plant tissue, frozen plant tissue, and processed planttissue; seeds and plant parts processed for animal feed for terrestrialand/or aquatic animal consumption, oil, meal, flour, flakes, bran,fiber, paper, tea, coffee, silage, crushed of whole grain, and any otherfood for human or animal consumption; biomasses and fuel products; andraw material in industry.

Collection of seeds: In the context of the present disclosure acollection of seeds is a grouping of seeds mainly containing similarkind of seeds, for example hybrid seeds of the disclosure, but that mayalso contain, mixed together with this first kind of seeds, a second,different kind of seeds, of one of the inbred parent lines. A commercialbag of hybrid seeds of the disclosure and containing also the inbredparental line seeds would be, for example such a collection of seeds.

Determinate plant: A determinate plant will grow to a fixed number ofnodes while an indeterminate plant continues to grow during the season.

Emergence: The rate that the seed germinates and sprouts out of theground.

Enhanced nutritional quality: The nutritional quality of the bean of thepresent disclosure can be enhanced by the introduction of several traitscomprising a higher vitamins, protein content in the pod, morebioavailable forms of vitamins and such, richer green color etc.

Essentially all of the physiological and morphological characteristics:A plant having essentially all of the physiological and morphologicalcharacteristics means a plant having all of the physiological andmorphological characteristics of a plant of the present disclosure,except for example, additional traits and/or mutations which do notmaterially affect the plant of the present disclosure, or, a desiredcharacteristic(s), which can be indirectly obtained from another plantpossessing at least one single locus conversion via a conventionalbreeding program (such as backcross breeding) or directly obtained byintroduction of at least one single locus conversion via New BreedingTechniques. In some embodiments, one of the non-limiting examples for aplant having (and/or comprising) essentially all of the physiologicaland morphological characteristics shall be a plant having all of thephysiological and morphological characteristics of a plant of thepresent disclosure other than desired, additionaltrait(s)/characteristic(s) conferred by a single locus conversionincluding, but not limited to, a converted or modified gene.

Field holding ability: A bean plant that has field holding ability meansa plant having pods that remain smooth and retain their color even afterthe seed is almost fully developed.

Immunity to disease(s) and or insect(s): A bean plant which is notsubject to attack or infection by specific disease(s) and or insect(s)is considered immune.

Intermediate resistance to disease(s) and or insect(s): A bean plantthat restricts the growth and development of specific disease(s) and orinsect(s), but may exhibit a greater range of symptoms or damagecompared to resistant plants. Intermediate resistant plants will usuallyshow less severe symptoms or damage than susceptible plant varietieswhen grown under similar environmental conditions and/or specificdisease(s) and or insect(s) pressure, but may have heavy damage underheavy pressure. Intermediate resistant bean plants are not immune to thedisease(s) and or insect(s).

New Breeding Techniques: New breeding techniques (NBTs) are said ofvarious new technologies developed and/or used to create newcharacteristics in plants through genetic variation, the aim beingtargeted mutagenesis, that is targeted introduction of new genes or genesilencing. The following breeding techniques are within the scope ofNBTs: targeted sequence changes facilitated through the use of Zincfinger nuclease (ZFN) technology (ZFN-1, ZFN-2 and ZFN-3, see U.S. Pat.No. 9,145,565, incorporated by reference in its entirety),Oligonucleotide directed mutagenesis (ODM, a.k.a., site-directedmutagenesis), Cisgenesis and intragenesis, epigenetic approaches such asRNA-dependent DNA methylation (RdDM, which does not necessarily changenucleotide sequence but can change the biological activity of thesequence), Grafting (on GM rootstock), Reverse breeding,Agro-infiltration for transient gene expression (agro-infiltration“sensu stricto”, agro-inoculation, floral dip), genome editing withendonucleases such as chemical nucleases, meganucleases, ZFNs, andTranscription Activator-Like Effector Nucleases (TALENs, see U.S. Pat.Nos. 8,586,363 and 9,181,535, incorporated by reference in theirentireties), the CRISPR/Cas system (using such as Cas9, Cas12a/Cpf1,Cas13/C2c2, CasX and CasY; also see U.S. Pat. Nos. 8,697,359; 8,771,945;8,795,965; 8,865,406; 8,871,445; 8,889,356; 8,895,308; 8,906,616;8,932,814; 8,945,839; 8,993,233; and 8,999,641, which are all herebyincorporated by reference), engineered meganuclease, engineered homingendonucleases, DNA guided genome editing (Gao et al., NatureBiotechnology (2016), doi: 10.1038/nbt.3547, incorporated by referencein its entirety), and Synthetic genomics. A major part of today'stargeted genome editing, another designation for New BreedingTechniques, is the applications to induce a DNA double strand break(DSB) at a selected location in the genome where the modification isintended. Directed repair of the DSB allows for targeted genome editing.Such applications can be utilized to generate mutations (e.g., targetedmutations or precise native gene editing) as well as precise insertionof genes (e.g., cisgenes, intragenes, or transgenes). The applicationsleading to mutations are often identified as site-directed nuclease(SDN) technology, such as SDN1, SDN2 and SDN3. For SDN1, the outcome isa targeted, non-specific genetic deletion mutation: the position of theDNA DSB is precisely selected, but the DNA repair by the host cell israndom and results in small nucleotide deletions, additions orsubstitutions. For SDN2, a SDN is used to generate a targeted DSB and aDNA repair template (a short DNA sequence identical to the targeted DSBDNA sequence except for one or a few nucleotide changes) is used torepair the DSB: this results in a targeted and predetermined pointmutation in the desired gene of interest. As to the SDN3, the SDN isused along with a DNA repair template that contains new DNA sequence(e.g. gene). The outcome of the technology would be the integration ofthat DNA sequence into the plant genome. The most likely applicationillustrating the use of SDN3 would be the insertion of cisgenic,intragenic, or transgenic expression cassettes at a selected genomelocation. A complete description of each of these techniques can befound in the report made by the Joint Research Center (JRC) Institutefor Prospective Technological Studies of the European Commission in 2011and titled “New plant breeding techniques—State-of-the-art and prospectsfor commercial development”, which is incorporated by reference in itsentirety.

Machine harvestable bush: A machine harvestable bush means a bean plantthat stands with pods off the ground. The pods can be removed by amachine from the plant without leaves and other plant parts.

Maturity or Relative Maturity: A maturity under 53 days is consideredearly while maturity between 54-59 days is considered average or mediumand maturity of 60 or more days would be late.

Maturity date: Plants are considered mature when the pods have reachedtheir maximum allowable seed size and sieve size for the specific useintended. This can vary for each end user, e.g., processing at differentstages of maturity would be required for different types of consumerbeans, such as “whole pack,” “cut,” or “french style.” The number ofdays is calculated from a relative planting date which depends on daylength, heat units, and other environmental factors.

Plant adaptability: A plant having good plant adaptability means a plantthat will perform well in different growing conditions and seasons.

Plant architecture: Plant architecture is the shape of the overall plantwhich can be tall narrow, short-wide, medium height, and/or mediumwidth.

Plant cell: As used herein, the term “plant cell” includes plant cellswhether isolated, in tissue culture, or incorporated in a plant or plantpart.

Plant habit: A plant can be erect (upright) to sprawling on the ground.

Plant height: Plant height is taken from the top of the soil to the topnode of the plant and is measured in centimeters or inches.

Plant part: As used herein, the term “plant part”, “part thereof” or“parts thereof” includes plant cells, plant protoplasts, plant celltissue cultures from which bean plants can be regenerated, plant calli,plant clumps and plant cells that are intact in plants or parts ofplants, such as pods, beans, embryos, pollens, ovules, flowers, seeds,fruits, rootstocks, scions, stems, roots, anthers, pistils, root tips,leaves, meristematic cells, axillary buds, hypocotyls, cotyledons,ovaries, seed coats, endosperms and the like. In some embodiments, theplant part at least comprises at least one cell of said plant. In someembodiments, the plant part is further defined as a pollen, a meristem,a cell or an ovule. In some embodiments, a plant regenerated from theplant part has all of the phenotypic and morphological characteristicsof a bean plant of the present disclosure, including but not limited toas determined at the 5% significance level when grown in the sameenvironmental conditions.

Pod set height: The pod set height is the location of the pods withinthe plant. The pods can be high (near the top), low (near the bottom),or medium (in the middle) of the plant.

Quantitative Trait Loci (QTL): Quantitative trait loci refer to geneticloci that control to some degree numerically representable traits thatare usually continuously distributed.

Regeneration: Regeneration refers to the development of a plant fromtissue culture.

Resistance to disease(s) and or insect(s): A bean plant that restrictsthe growth and development of specific disease(s) and or insect(s) undernormal disease(s) and or insect(s) attack pressure when compared tosusceptible plants. These bean plants can exhibit some symptoms ordamage under heavy disease(s) and or insect(s) pressure. Resistant beanplants are not immune to the disease(s) and or insect(s).

RHS: RHS refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Hort. Society Enterprise Ltd. RHS Garden; Wisley,Woking, Surrey GU236QB, UK.

Rootstock: A rootstock is the lower part of a plant capable of receivinga scion in a grafting process.

Scion: A scion is the higher part of a plant capable of being graftedonto a rootstock in a grafting process.

Sieve size (sv): Sieve size 1 means pods that fall through a sievegrader which culls out pod diameters of 4.76 mm through 5.76 mm. Sievesize 2 means pods that fall through a sieve grader which culls out poddiameters of 5.76 mm through 7.34 mm. Sieve size 3 means pods that fallthrough a sieve grader which culls out pod diameters of 7.34 mm through8.34 mm. Sieve size 4 means pods that fall through a sieve grader whichculls out pod diameters of 8.34 mm through 9.53 mm. Sieve size 5 meanspods that fall through a sieve grader which culls out pod diameters of9.53 mm through 10.72 mm. Sieve size 6 means pods that fall through asieve grader that will cull out pod diameters of 10.72 mm or larger.

Single locus converted (conversion): Single locus converted (conversion)plants refer to plants which are developed by a plant breeding techniquecalled backcrossing, wherein essentially all of the desiredmorphological and physiological characteristics of a plant are recoveredin addition to a single locus transferred into the plant via thebackcrossing technique or via genetic engineering. A single locusconverted plant can also be referred to a plant with a single locusconversion obtained though simultaneous and/or artificially inducedmutagenesis or through the use of New Breeding Techniques described inthe present disclosure. In some embodiments, the single locus convertedplant has essentially all of the desired morphological and physiologicalcharacteristics of the original variety in addition to a single locusconverted by spontaneous and/or artificially induced mutations, which isintroduced and/or transferred into the plant by the plant breedingtechniques such as backcrossing. In other embodiments, the single locusconverted plant has essentially all of the desired morphological andphysiological characteristics of the original variety in addition to asingle locus, gene or nucleotide sequence(s) converted, mutated,modified or engineered through the New Breeding Techniques taughtherein. In the present disclosure, single locus converted (conversion)can be interchangeably referred to single gene converted (conversion).

Slow seed development: Beans having slow seed development develop seedslowly even after the pods are full sized. This characteristic gives tothe cultivar its field holding ability.

Susceptible to disease(s) and or insect(s): A bean plant that issusceptible to disease(s) and or insect(s) is defined as a bean plantthat has the inability to restrict the growth and development ofspecific disease(s) and or insect(s). Plants that are susceptible willshow damage when infected and are more likely to have heavy damage undermoderate levels of specific disease(s) and or insect(s).

Tolerance to abiotic stresses: A bean plant that is tolerant to abioticstresses has the ability to endure abiotic stress without seriousconsequences for growth, appearance and yield.

Uniformity: Uniformity, as used herein, describes the similarity betweenplants or plant characteristics which can be a described by qualitativeor quantitative measurements.

Variety: A plant variety as used by one skilled in the art of plantbreeding means a plant grouping within a single botanical taxon of thelowest known rank which can be defined by the expression of thecharacteristics resulting from a given genotype or combination ofphenotypes, distinguished from any other plant grouping by theexpression of at least one of the said characteristics and considered asa unit with regard to its suitability for being propagated unchanged(International convention for the protection of new varieties ofplants). The term “bean variety” can be interchangeably used with “beancultivar” or “bean plant” in the present application.

There are numerous steps in the development of any novel, desirableplant germplasm. Plant breeding begins with the analysis and definitionof problems and weaknesses of the current germplasm, the establishmentof program goals, and the definition of specific breeding objectives.The next step is selection of germplasm that possesses the traits tomeet the program goals. The goal is to combine in a single variety orhybrid an improved combination of desirable traits from the parentalgermplasm.

Bean Plants

Phaseolus vulgaris, also known as the common bean and French bean, is aherbaceous annual plant grown worldwide for its edible dry seeds orunripe fruit (both commonly called beans). The main categories of commonbeans, on the basis of use, are dry beans (seeds harvested at completematurity), snap beans (tender pods with reduced fiber harvested beforethe seed development phase) and shell (shelled) beans (seeds harvestedat physiological maturity). Its leaf is also occasionally used as avegetable and the straw as fodder. Its botanical classification, alongwith other Phaseolus species, is as a member of the legume familyFabaceae. Like most members of this family, common beans acquire thenitrogen they require through an association with rhizobia, which arenitrogen-fixing bacteria.

Wild members of the species have a climbing habit, but many cultivarsare classified either as bush beans or dwarf beans, or as pole beans orclimbing beans, depending on their style of growth. These include thekidney bean, the navy bean, the pinto bean, and the wax bean. The othermajor types of commercially grown bean are the runner bean (Phaseoluscoccineus) and the broad bean (Vicia faba).

There are numerous varieties of P. vulgaris, including many commongarden types (i.e., garden bean) such as pole, snap, string, and bushbeans. It is called French bean, haricot bean, or kidney bean in variouscountries; in the United States, however, kidney bean refers to aspecific type that is definitely kidney-shaped and is red, dark red, orwhite. Green beans, anasazi beans, navy beans, black beans, northernbeans, kidney beans, pinto beans, and cannellini beans are all varietiesof the species. Some varieties of the common bean are grown only for thedry seeds, some only for the edible immature pods, and others for theseeds, either immature or mature.

Many well-known bean cultivars and varieties belong to Phaseolusvulgaris. An exemplary list of the bean cultivars and varieties, but arenot limited to, are as follows: Anasazi bean, appaloosa bean, blackturtle bean, calypso bean, cranberry bean, dragon tongue bean, flageoletbean, kidney bean (a.k.a. red bean), pea bean, pink bean, pinto bean,rattlesnake bean, white bean (a.k.a. navy bean or haricot bean'including cannellini), yellow bean (a.k.a. Sinaloa Azufrado, Sulphur,Mayacoba, Peruano, Canary), and tongue of fire bean (a.k.a. Horto)

In bean, these important traits may include increased fresh pod yield,higher seed yield, improved color, resistance to diseases and insects,better stems and roots, tolerance to drought and heat, better standingability in the field, better uniformity, and better agronomic quality.With mechanical harvesting of many crops, uniformity of plantcharacteristics such as germination and stand establishment, growthrate, maturity and plant height is important.

In some embodiments, particularly desirable traits that may beincorporated by this disclosure are improved resistance to differentviral, fungal, and bacterial pathogens. Important diseases include butare not limited to fungi such as Uromyces appendiculatus (rust),Colletotrichum lindemuthianum (anthracnose), virus such as BCMV (beancommon mosaic virus), BCTV (bean curly top virus), bacteria such asPseudomonas (Pseudomonas savastanoi pv. Phaseolicola (halo blight),Pseudomonas syringae pv. Syringae (bacterial brown spot)) or Xanthomonas(Xanthomonas axonopodis pv. Phaseoli (common blight). Improvedresistance to insect pests is another desirable trait that may beincorporated into new garden plants developed by this disclosure.

Bean Breeding

The goal of bean breeding is to develop new, unique and superior beanplant and hybrids. The breeder initially selects and crosses two or moreparental lines, followed by repeated selfing and selection, producingmany new genetic combinations. Another method used to develop new,unique and superior bean plant occurs when the breeder selects andcrosses two or more parental lines followed by haploid induction andchromosome doubling that result in the development of dihaploidcultivars. The breeder can theoretically generate billions of differentgenetic combinations via crossing, selfing and mutations and the same istrue for the utilization of the dihaploid breeding method.

Each year, the plant breeder selects the germplasm to advance to thenext generation. This germplasm is grown under unique and differentgeographical, climatic and soil conditions, and further selections arethen made, during and at the end of the growing season. The cultivarsdeveloped are unpredictable. This unpredictability is because thebreeder's selection occurs in unique environments, with no control atthe DNA level (using conventional breeding procedures or dihaploidbreeding procedures), and with millions of different possible geneticcombinations being generated. A breeder of ordinary skill in the artcannot predict the final resulting cultivars he develops, exceptpossibly in a very gross and general fashion. This unpredictabilityresults in the expenditure of large research monies to develop superiornew bean cultivars.

The development of commercial bean cultivar requires the development andselection of bean plants, the crossing of these plants, and theevaluation of the crosses.

Pedigree breeding and recurrent selection breeding methods are used todevelop cultivars from breeding populations. Breeding programs combinedesirable traits from two or more cultivars or various broad-basedsources into breeding pools from which cultivars are developed byselfing and selection of desired phenotypes or through the dihaploidbreeding method followed by the selection of desired phenotypes. The newcultivars are evaluated to determine which have commercial potential.

Choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially (e.g., F₁ hybrid cultivar, purelinecultivar, etc.). For highly heritable traits, a choice of superiorindividual plants evaluated at a single location will be effective,whereas for traits with low heritability, selection should be based onmean values obtained from replicated evaluations of families of relatedplants. Popular selection methods commonly include pedigree selection,modified pedigree selection, mass selection, recurrent selection, andbackcross breeding.

i. Pedigree Selection

Pedigree breeding is used commonly for the improvement ofself-pollinating crops or inbred lines of cross-pollinating crops. Twoparents possessing favorable, complementary traits are crossed toproduce an F₁. An F₂ population is produced by selfing one or severalF₁s or by intercrossing two F₁s (sib mating). The dihaploid breedingmethod could also be used. Selection of the best individuals is usuallybegun in the F₂ population; then, beginning in the F₃, the bestindividuals in the best families are selected. Replicated testing offamilies, or hybrid combinations involving individuals of thesefamilies, often follows in the F₄ generation to improve theeffectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines are tested for potentialrelease of new cultivars. Similarly, the development of new cultivarsthrough the dihaploid system requires the selection of the cultivarsfollowed by two to five years of testing in replicated plots.

The single-seed descent procedure in the strict sense refers to plantinga segregating population, harvesting a sample of one seed per plant, andusing the one-seed sample to plant the next generation. When thepopulation has been advanced from the F2 to the desired level ofinbreeding, the plants from which lines are derived will each trace todifferent F2 individuals. The number of plants in a population declineseach generation due to failure of some seeds to germinate or some plantsto produce at least one seed. As a result, not all of the F2 plantsoriginally sampled in the population will be represented by a progenywhen generation advance is completed.

In a multiple-seed procedure, breeders commonly harvest one or morefruit containing seed from each plant in a population and blend themtogether to form a bulk seed lot. Part of the bulked seed is used toplant the next generation and part is put in reserve. The procedure hasbeen referred to as modified single-seed descent or the bulk technique.

The multiple-seed procedure has been used to save labor at harvest. Itis considerably faster than removing one seed from each fruit by handfor the single seed procedure. The multiple-seed procedure also makes itpossible to plant the same number of seeds of a population eachgeneration of inbreeding. Enough seeds are harvested to make up forthose plants that did not germinate or produce seed.

Descriptions of other breeding methods that are commonly used fordifferent traits and crops can be found in one of several referencebooks (e.g., R. W. Allard, 1960, Principles of Plant Breeding, JohnWiley and Son, pp. 115-161; N.W. Simmonds, 1979, Principles of CropImprovement, Longman Group Limited; W. R. Fehr, 1987, Principles of CropDevelopment, Macmillan Publishing Co.; N. F. Jensen, 1988, PlantBreeding Methodology, John Wiley & Sons).

ii. Backcross Breeding

Backcross breeding has been used to transfer genes for a simplyinherited, highly heritable trait into a desirable homozygous cultivaror inbred line which is the recurrent parent. The source of the trait tobe transferred is called the donor parent. The resulting plant isexpected to have the attributes of the recurrent parent (e.g., cultivar)and the desirable trait transferred from the donor parent. After theinitial cross, individuals possessing the phenotype of the donor parentare selected and repeatedly crossed (backcrossed) to the recurrentparent. The resulting plant is expected to have the attributes of therecurrent parent (e.g., cultivar) and the desirable trait transferredfrom the donor parent.

When the term bean plant is used in the context of the presentdisclosure, this also includes any bean plant where one or more desiredtraits have been introduced through backcrossing methods, whether suchtrait is from a naturally occurring mutation(s), an artificially-inducedmutation(s), a gene or a nucleotide sequence modified by the use of NewBreeding Techniques. Backcrossing methods can be used with the presentdisclosure to improve or introduce one or more characteristic into thebean plant of the present disclosure. The term “backcrossing” as usedherein refers to the repeated crossing of a hybrid progeny back to therecurrent parent, i.e., backcrossing one, two, three, four, five, six,seven, eight, nine, or more times to the recurrent parent. The parentalbean plant which contributes the gene or the genes for the desiredcharacteristic is termed the nonrecurrent or donor parent. Thisterminology refers to the fact that the nonrecurrent parent is used onetime in the backcross protocol and therefore does not recur. Theparental bean plant to which the gene or genes from the nonrecurrentparent are transferred is known as the recurrent parent as it is usedfor several rounds in the backcrossing protocol.

In a typical backcross protocol, the original cultivar of interest(recurrent parent) is crossed to or by a second cultivar (nonrecurrentparent) that carries the gene or genes of interest to be transferred.The resulting progeny from this cross are then crossed again to or bythe recurrent parent and the process is repeated until a bean plant isobtained wherein all of the desired morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, generally determined at a 5% significance level when grown in thesame environmental conditions, in addition to the gene or genestransferred from the nonrecurrent parent. It has to be noted that some,one, two, three or more, self-pollination and growing of populationmight be included between two successive backcrosses. Indeed, anappropriate selection in the population produced by theself-pollination, i.e., selection for the desired trait andphysiological and morphological characteristics of the recurrent parentmight be equivalent to one, two or even three additional backcrosses ina continuous series without rigorous selection, saving then time, moneyand effort to the breeder. A non-limiting example of such a protocolwould be the following: a) the first generation F1 produced by the crossof the recurrent parent A by the donor parent B is backcrossed to parentA, b) selection is practiced for the plants having the desired trait ofparent B, c) selected plant are self-pollinated to produce a populationof plants where selection is practiced for the plants having the desiredtrait of parent B and physiological and morphological characteristics ofparent A, d) the selected plants are backcrossed one, two, three, four,five, six, seven, eight, nine, or more times to parent A to produceselected backcross progeny plants comprising the desired trait of parentB and the physiological and morphological characteristics of parent A.Step (c) may or may not be repeated and included between the backcrossesof step (d).

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute one or more trait(s) or characteristic(s) in theoriginal inbred parental line in order to find it then in the hybridmade thereof. To accomplish this, a gene or genes of the recurrentinbred is modified or substituted with the desired gene or genes fromthe nonrecurrent parent, while retaining essentially all of the rest ofthe desired genetic, and therefore the desired physiological andmorphological, constitution of the original inbred. The choice of theparticular nonrecurrent parent will depend on the purpose of thebackcross; one of the major purposes is to add some commerciallydesirable, agronomically important trait(s) to the plant. The exactbackcrossing protocol will depend on the characteristic(s) or trait(s)being altered to determine an appropriate testing protocol. Althoughbackcrossing methods are simplified when the characteristic beingtransferred is a single gene and dominant allele, multiple genes andrecessive allele(s) may also be transferred and therefore, backcrossbreeding is by no means restricted to character(s) governed by one or afew genes. In fact the number of genes might be less important that theidentification of the character(s) in the segregating population. Inthis instance it may then be necessary to introduce a test of theprogeny to determine if the desired characteristic(s) has beensuccessfully transferred. Such tests encompass visual inspection, simplecrossing, but also follow up of the characteristic(s) throughgenetically associated markers and molecular assisted breeding tools.For example, selection of progeny containing the transferred trait isdone by direct selection, visual inspection for a trait associated witha dominant allele, while the selection of progeny for a trait that istransferred via a recessive allele, require selfing the progeny or usingmolecular markers to determine which plant carry the recessiveallele(s).

Many single gene traits have been identified that are not regularlyselected for in the development of a new parental inbred of a hybridbean plant according to the disclosure but that can be improved bybackcrossing techniques. Examples of these traits include but are notlimited to herbicide resistance (such as bar or pat genes), resistancefor bacterial, fungal, or viral disease (such as gene I used for BCMVresistance), insect resistance, enhanced nutritional quality (such as 2salbumin gene), industrial usage, agronomic qualities (such as the“persistent green gene”), yield stability, and yield enhancement. Manysingle gene traits have been identified that are not regularly selectedfor in the development of a new line but that can be improved bybackcrossing techniques. These genes are generally inherited through thenucleus. Some other single gene traits are described in U.S. Pat. Nos.5,777,196, 5,948,957, and 5,969,212, the disclosures of which arespecifically hereby incorporated by reference.

The backcross breeding method provides a precise way of improvingvarieties that excel in a large number of attributes but are deficientin a few characteristics. (Page 150 of the Pr. R. W. Allard's 1960 book,published by John Wiley & Sons, Inc, Principles of Plant Breeding). Themethod makes use of a series of backcrosses to the variety to beimproved during which the character or the characters in whichimprovement is sought is maintained by selection. At the end of thebackcrossing the gene or genes being transferred unlike all other genes,will be heterozygous. Selfing after the last backcross produceshomozygosity for this gene pair(s) and, coupled with selection, willresult in a parental line of a hybrid variety with exactly oressentially the same adaptation, yielding ability and qualitycharacteristics of the recurrent parent but superior to that parent inthe particular characteristic(s) for which the improvement program wasundertaken. Therefore, this method provides the plant breeder with ahigh degree of genetic control of his work.

The method is scientifically exact because the morphological andagricultural features of the improved variety could be described inadvance and because the same variety could, if it were desired, be breda second time by retracing the same steps (Briggs, “Breeding wheatsresistant to bunt by the backcross method”, 1930 Jour. Amer. Soc.Agron., 22: 289-244).

Backcrossing is a powerful mechanism for achieving homozygosity and anypopulation obtained by backcrossing must rapidly converge on thegenotype of the recurrent parent. When backcrossing is made the basis ofa plant breeding program, the genotype of the recurrent parent will bemodified only with regards to genes being transferred, which aremaintained in the population by selection.

Successful backcrosses are, for example, the transfer of stem rustresistance from ‘Hope’ wheat to ‘Bart wheat’ and even pursuing thebackcrosses with the transfer of bunt resistance to create ‘Bart 38’,having both resistances. Also highlighted by Allard is the successfultransfer of mildew, leaf spot and wilt resistances in California Commonalfalfa to create ‘Caliverde’. This new ‘Caliverde’ variety producedthrough the backcross process is indistinguishable from CaliforniaCommon except for its resistance to the three named diseases.

One of the advantages of the backcross method is that the breedingprogram can be carried out in almost every environment that will allowthe development of the character being transferred.

The backcross technique is not only desirable when breeding for diseaseresistance but also for the adjustment of morphological characters,colour characteristics and simply inherited quantitative characters suchas earliness, plant height and seed size and shape.

iii. Open-Pollinated Populations

The improvement of open-pollinated populations of such crops as rye,maize, sugar beets, herbage grasses, legumes such as alfalfa and clover,and tropical tree crops such as cacao, coconuts, oil palm and somerubber, depends essentially upon changing gene-frequencies towardsfixation of favorable alleles while maintaining a high (but far frommaximal) degree of heterozygosity.

Uniformity in such populations is impossible and trueness-to-type in anopen-pollinated variety is a statistical feature of the population as awhole, not a characteristic of individual plants. Thus, theheterogeneity of open-pollinated populations contrasts with thehomogeneity (or virtually so) of inbred lines, clones and hybrids.

Population improvement methods fall naturally into two groups, thosebased on purely phenotypic selection, normally called mass selection,and those based on selection with progeny testing. Interpopulationimprovement utilizes the concept of open breeding populations; allowinggenes to flow from one population to another. Plants in one population(cultivar, strain, ecotype, or any germplasm source) are crossed eithernaturally (e.g., by wind) or by hand or by bees (commonly Apis melliferaL. or Megachile rotundata F.) with plants from other populations.Selection is applied to improve one (or sometimes both) population(s) byisolating plants with desirable traits from both sources.

There are basically two primary methods of open-pollinated populationimprovement.

First, there is the situation in which a population is changed en masseby a chosen selection procedure. The outcome is an improved populationthat is indefinitely propagable by random-mating within itself inisolation.

Second, the synthetic variety attains the same end result as populationimprovement, but is not itself propagable as such; it has to bereconstructed from parental lines or clones. These plant breedingprocedures for improving open-pollinated populations are well known tothose skilled in the art and comprehensive reviews of breedingprocedures routinely used for improving cross-pollinated plants areprovided in numerous texts and articles, including: Allard, Principlesof Plant Breeding, John Wiley & Sons, Inc. (1960); Simmonds, Principlesof Crop Improvement, Longman Group Limited (1979); Hallauer and Miranda,Quantitative Genetics in Maize Breeding, Iowa State University Press(1981); and, Jensen, Plant Breeding Methodology, John Wiley & Sons, Inc.(1988).

A) Mass Selection

Mass and recurrent selections can be used to improve populations ofeither self- or cross-pollinating crops. A genetically variablepopulation of heterozygous individuals is either identified or createdby intercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued. In massselection, desirable individual plants are chosen, harvested, and theseed composited without progeny testing to produce the followinggeneration. Since selection is based on the maternal parent only, andthere is no control over pollination, mass selection amounts to a formof random mating with selection. As stated above, the purpose of massselection is to increase the proportion of superior genotypes in thepopulation.

B) Synthetics

A synthetic variety is produced by crossing inter se a number ofgenotypes selected for good combining ability in all possible hybridcombinations, with subsequent maintenance of the variety by openpollination. Whether parents are (more or less inbred) seed-propagatedlines, as in some sugar beet and beans (Vicia) or clones, as in herbagegrasses, clovers and alfalfa, makes no difference in principle. Parentsare selected on general combining ability, sometimes by test crosses ortoperosses, more generally by polycrosses. Parental seed lines may bedeliberately inbred (e.g. by selfing or sib crossing). However, even ifthe parents are not deliberately inbred, selection within lines duringline maintenance will ensure that some inbreeding occurs. Clonal parentswill, of course, remain unchanged and highly heterozygous.

Whether a synthetic can go straight from the parental seed productionplot to the farmer or must first undergo one or more cycles ofmultiplication depends on seed production and the scale of demand forseed. In practice, grasses and clovers are generally multiplied once ortwice and are thus considerably removed from the original synthetic.

While mass selection is sometimes used, progeny testing is generallypreferred for polycrosses, because of their operational simplicity andobvious relevance to the objective, namely exploitation of generalcombining ability in a synthetic.

The number of parental lines or clones that enters a synthetic varieswidely. In practice, numbers of parental lines range from 10 to severalhundred, with 100-200 being the average. Broad based synthetics formedfrom 100 or more clones would be expected to be more stable during seedmultiplication than narrow based synthetics.

iv. Hybrids

A hybrid is an individual plant resulting from a cross between parentsof differing genotypes. Commercial hybrids are now used extensively inmany crops, including corn (maize), sorghum, sugar beet, sunflower andbroccoli. Hybrids can be formed in a number of different ways, includingby crossing two parents directly (single cross hybrids), by crossing asingle cross hybrid with another parent (three-way or triple crosshybrids), or by crossing two different hybrids (four-way or double crosshybrids).

Strictly speaking, most individuals in an out breeding (i.e.,open-pollinated) population are hybrids, but the term is usuallyreserved for cases in which the parents are individuals whose genomesare sufficiently distinct for them to be recognized as different speciesor subspecies. Hybrids may be fertile or sterile depending onqualitative and/or quantitative differences in the genomes of the twoparents. Heterosis, or hybrid vigor, is usually associated withincreased heterozygosity that results in increased vigor of growth,survival, and fertility of hybrids as compared with the parental linesthat were used to form the hybrid. Maximum heterosis is usually achievedby crossing two genetically different, highly inbred lines.

Once the inbreds that give the best hybrid performance have beenidentified, the hybrid seed can be reproduced indefinitely as long asthe homogeneity of the inbred parent is maintained. A single-crosshybrid is produced when two inbred lines are crossed to produce the F1progeny. A double-cross hybrid is produced from four inbred linescrossed in pairs (A×B and C×D) and then the two F1 hybrids are crossedagain (A×B)×(C×D). Much of the hybrid vigor and uniformity exhibited byF1 hybrids is lost in the next generation (F2). Consequently, seed fromF2 hybrid varieties is not used for planting stock. The production ofhybrids is a well-developed industry, involving the isolated productionof both the parental lines and the hybrids which result from crossingthose lines. For a detailed discussion of the hybrid production process,see, e.g., Wright, Commercial Hybrid Seed Production 8:161-176, InHybridization of Crop Plants.

v. Bulk Segregation Analysis (BSA)

BSA, a.k.a. bulked segregation analysis, or bulk segregant analysis, isa method described by Michelmore et al. (Michelmore et al., 1991,Identification of markers linked to disease-resistance genes by bulkedsegregant analysis: a rapid method to detect markers in specific genomicregions by using segregating populations. Proceedings of the NationalAcademy of Sciences, USA, 99:9828-9832) and Quarrie et al. (Quarrie etal., 1999, Journal of Experimental Botany, 50(337): 1299-1306).

For BSA of a trait of interest, parental lines with certain differentphenotypes are chosen and crossed to generate F2, doubled haploid orrecombinant inbred populations with QTL analysis. The population is thenphenotyped to identify individual plants or lines having high or lowexpression of the trait. Two DNA bulks are prepared, one from theindividuals having one phenotype (e.g., resistant to virus), and theother from the individuals having reversed phenotype (e.g., susceptibleto virus), and analyzed for allele frequency with molecular markers.Only a few individuals are required in each bulk (e.g., 10 plants each)if the markers are dominant (e.g., RAPDs). More individuals are neededwhen markers are co-dominant (e.g., RFLPs, SNPs or SSRs). Markers linkedto the phenotype can be identified and used for breeding or QTL mapping.

vi. Hand-Pollination Method

Hand pollination describes the crossing of plants via the deliberatefertilization of female ovules with pollen from a desired male parentplant. In some embodiments the donor or recipient female parent and thedonor or recipient male parent line are planted in the same field. Insome embodiments the donor or recipient female parent and the donor orrecipient male parent line are planted in the same greenhouse. Theinbred male parent can be planted earlier than the female parent toensure adequate pollen supply at the pollination time. In someembodiments, the male parent and female parent can be planted at a ratioof 1 male parent to 4-10 female parents. Pollination is started when thefemale parent flower is ready to be fertilized. Female flower buds thatare ready to open in the following days are identified, covered withpaper cups or small paper bags that prevent bee or any other insect fromvisiting the female flowers, and marked with any kind of material thatcan be easily seen the next morning. The male flowers of the male parentare collected in the early morning before they are open and visited bypollinating insects. The covered female flowers of the female parent,which have opened, are un-covered and pollinated with the collectedfresh male flowers of the male parent, starting as soon as the maleflower sheds pollen. The pollinated female flowers are again coveredafter pollination to prevent bees and any other insects visit. Thepollinated female flowers are also marked. The marked flowers areharvested. In some embodiments, the male pollen used for fertilizationhas been previously collected and stored.

vii. Bee-Pollination Method

Using the bee-pollination method, the parent plants are usually plantedwithin close proximity. In some embodiments more female plants areplanted to allow for a greater production of seed. Insects are placed inthe field or greenhouses for transfer of pollen from the male parent tothe female flowers of the female parent.

viii. Targeting Induced Local Lesions in Genomes (TILLING)

Breeding schemes of the present application can include crosses withTILLING® plant cultivars. TILLING® is a method in molecular biology thatallows directed identification of mutations in a specific gene. TILLING®was introduced in 2000, using the model plant Arabidopsis thaliana.TILLING® has since been used as a reverse genetics method in otherorganisms such as zebrafish, corn, wheat, rice, soybean, tomato andlettuce.

The method combines a standard and efficient technique of mutagenesiswith a chemical mutagen (e.g., Ethyl methanesulfonate (EMS)) with asensitive DNA screening-technique that identifies single base mutations(also called point mutations) in a target gene. EcoTILLING is a methodthat uses TILLING® techniques to look for natural mutations inindividuals, usually for population genetics analysis (see Comai, etal., 2003 The Plant Journal 37, 778-786; Gilchrist et al. 2006 Mol.Ecol. 15, 1367-1378; Mejlhede et al. 2006 Plant Breeding 125, 461-467;Nieto et al. 2007 BMC Plant Biology 7, 34-42, each of which isincorporated by reference hereby for all purposes). DEcoTILLING is amodification of TILLING® and EcoTILLING which uses an inexpensive methodto identify fragments (Garvin et al., 2007, DEco-TILLING: An inexpensivemethod for SNP discovery that reduces ascertainment bias. MolecularEcology Notes 7, 735-746).

The TILLING® method relies on the formation of heteroduplexes that areformed when multiple alleles (which could be from a heterozygote or apool of multiple homozygotes and heterozygotes) are amplified in a PCR,heated, and then slowly cooled. A “bubble” forms at the mismatch of thetwo DNA strands (the induced mutation in TILLING® or the naturalmutation or SNP in EcoTILLING), which is then cleaved by single strandednucleases. The products are then separated by size on several differentplatforms.

Several TILLING® centers exists over the world that focus onagriculturally important species: UC Davis (USA), focusing on Rice;Purdue University (USA), focusing on Maize; University of BritishColumbia (CA), focusing on Brassica napus; John Innes Centre (UK),focusing on Brassica rapa; Fred Hutchinson Cancer Research, focusing onArabidopsis; Southern Illinois University (USA), focusing on Soybean;John Innes Centre (UK), focusing on Lotus and Medicago; and INRA(France), focusing on Pea and Tomato.

More detailed description on methods and compositions on TILLING® can befound in U.S. Pat. No. 5,994,075, US 2004/0053236 A1, WO 2005/055704,and WO 2005/048692, each of which is hereby incorporated by referencefor all purposes.

Thus, in some embodiments, the breeding methods of the presentdisclosure include breeding with one or more TILLING plant lines withone or more identified mutations.

ix. Mutation Breeding

Mutation breeding is another method of introducing new traits into beanplants. Mutations that occur spontaneously or are artificially inducedcan be useful sources of variability for a plant breeder. The goal ofartificial mutagenesis is to increase the rate of mutation for a desiredcharacteristic. Mutation rates can be increased by many different meansor mutating agents including temperature, long-term seed storage, tissueculture conditions, radiation (such as X-rays, Gamma rays, neutrons,Beta radiation, or ultraviolet radiation), chemical mutagens (such asbase analogs like 5-bromo-uracil), antibiotics, alkylating agents (suchas sulfur mustards, nitrogen mustards, epoxides, ethyleneamines,sulfates, sulfonates, sulfones, or lactones), azide, hydroxylamine,nitrous acid or acridines. Once a desired trait is observed throughmutagenesis the trait may then be incorporated into existing germplasmby traditional breeding techniques. Details of mutation breeding can befound in W. R. Fehr, 1993, Principles of Cultivar Development, MacmillanPublishing Co.

New breeding techniques such as the ones involving the uses ofengineered nuclease to enhance the efficacy and precision of geneediting in combination with oligonucleotides including, but not limitedto Zinc Finger Nucleases (ZFN), TAL effector nucleases (TALENs) andclustered regularly interspaced short palindromic repeats(CRISPR)-associated endonuclease Cas9 (CRISPR-Cas9) using such as Cas9,Cas12a/Cpf1, Cas13/C2c2, CasX and CasY or oligonucleotide directedmutagenesis shall also be used to generate genetic variability andintroduce new traits into bean varieties.

x. Double Haploids and Chromosome Doubling

One way to obtain homozygous plants without the need to cross twoparental lines followed by a long selection of the segregating progeny,and/or multiple back-crossings is to produce haploids and then doublethe chromosomes to form doubled haploids. Haploid plants can occurspontaneously, or may be artificially induced via chemical treatments orby crossing plants with inducer lines (Seymour et al. 2012, PNAS vol109, pg 4227-4232; Zhang et al., 2008 Plant Cell Rep. Dec 27(12)1851-60). The production of haploid progeny can occur via a variety ofmechanisms which can affect the distribution of chromosomes duringgamete formation. The chromosome complements of haploids sometimesdouble spontaneously to produce homozygous doubled haploids (DHs).Mixoploids, which are plants which contain cells having differentploidies, can sometimes arise and may represent plants that areundergoing chromosome doubling so as to spontaneously produce doubledhaploid tissues, organs, shoots, floral parts or plants. Another commontechnique is to induce the formation of double haploid plants with achromosome doubling treatment such as colchicine (El-Hennawy et al.,2011 Vol 56, issue 2 pg 63-72; Doubled Haploid Production in Crop Plants2003 edited by Maluszynski ISBN 1-4020-1544-5). The production ofdoubled haploid plants yields highly uniform cultivars and is especiallydesirable as an alternative to sexual inbreeding of longer-generationcrops. By producing doubled haploid progeny, the number of possible genecombinations for inherited traits is more manageable. Thus, an efficientdoubled haploid technology can significantly reduce the time and thecost of inbred and cultivar development.

xi. Protoplast Fusion

In another method for breeding plants, protoplast fusion can also beused for the transfer of trait-conferring genomic material from a donorplant to a recipient plant. Protoplast fusion is an induced orspontaneous union, such as a somatic hybridization, between two or moreprotoplasts (cells of which the cell walls are removed by enzymatictreatment) to produce a single bi- or multi-nucleate cell. The fusedcell that may even be obtained with plant species that cannot beinterbred in nature is tissue cultured into a hybrid plant exhibitingthe desirable combination of traits.

xii. Embryo Rescue

Alternatively, embryo rescue may be employed in the transfer ofresistance-conferring genomic material from a donor plant to a recipientplant. Embryo rescue can be used as a procedure to isolate embryo's fromcrosses wherein plants fail to produce viable seed. In this process, thefertilized ovary or immature seed of a plant is tissue cultured tocreate new plants (see Pierik, 1999, In vitro culture of higher plants,Springer, ISBN 079235267x, 9780792352679, which is incorporated hereinby reference in its entirety).

Grafting

Grafting is a process that has been used for many years in crops. Butgrafting can be a tool in common bean breeding (Gurusamy et al. Can. J.Plant Sci. 90:299-304, 2010), which teaches that grafting shoots ofdifferent Phaseolus breeding materials onto compatible rootstocks cansuccessfully be used as a tool in bean breeding programs. Grafting maybe used to provide a certain level of resistance to telluric pathogenssuch as Phytophthora or to certain nematodes. Grafting is thereforeintended to prevent contact between the plant or variety to becultivated and the infested soil. The variety of interest used as thegraft or scion, optionally an F₁ hybrid, is grafted onto the resistantplant used as the rootstock. The resistant rootstock remains healthy andprovides, from the soils, the normal supply for the graft that itisolates from the diseases. In some recent developments, it has alsobeen shown that some rootstocks are also able to improve the agronomicvalue for the grafted plant and in particular the equilibrium betweenthe vegetative and generative development that are always difficult tobalance in bean cultivation.

Breeding Evaluation

Each breeding program can include a periodic, objective evaluation ofthe efficiency of the breeding procedure. Evaluation criteria varydepending on the goal and objectives, but should include gain fromselection per year based on comparisons to an appropriate standard,overall value of the advanced breeding lines, and number of successfulcultivars produced per unit of input (e.g., per year, per dollarexpended, etc.).

Promising advanced breeding lines are thoroughly tested per se and inhybrid combination and compared to appropriate standards in environmentsrepresentative of the commercial target area(s). The best lines arecandidates for use as parents in new commercial cultivars; those stilldeficient in a few traits may be used as parents to produce newpopulations for further selection or in a backcross program to improvethe parent lines for a specific trait.

In one embodiment, the plants are selected on the basis of one or morephenotypic traits. Skilled persons will readily appreciate that suchtraits include any observable characteristic of the plant, including forexample growth rate, vigor, plant health, maturity, branching, height,weight, total yield, color, taste, aroma, changes in the production ofone or more compounds by the plant (including for example, metabolites,proteins, drugs, carbohydrates, oils, and any other compounds).

A most difficult task is the identification of individuals that aregenetically superior, because for most traits the true genotypic valueis masked by other confounding plant traits or environmental factors.One method of identifying a superior plant is to observe its performancerelative to other experimental plants and to a widely grown standardcultivar. If a single observation is inconclusive, replicatedobservations provide a better estimate of its genetic worth.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current cultivars. In addition toshowing superior performance, there must be a demand for a new cultivarthat is compatible with industry standards or which creates a newmarket. The introduction of a new cultivar will incur additional coststo the seed producer, the grower, processor and consumer; for specialadvertising and marketing, altered seed and commercial productionpractices, and new product utilization. The testing preceding release ofa new cultivar should take into consideration research and developmentcosts as well as technical superiority of the final cultivar. Forseed-propagated cultivars, it must be feasible to produce seed easilyand economically.

It should be appreciated that in certain embodiments, plants may beselected based on the absence, suppression or inhibition of a certainfeature or trait (such as an undesirable feature or trait) as opposed tothe presence of a certain feature or trait (such as a desirable featureor trait).

Selecting plants based on genotypic information is also envisaged (forexample, including the pattern of plant gene expression, genotype, orpresence of genetic markers). Where the presence of one or more geneticmarker is assessed, the one or more marker may already be known and/orassociated with a particular characteristic of a plant; for example, amarker or markers may be associated with an increased growth rate ormetabolite profile. This information could be used in combination withassessment based on other characteristics in a method of the disclosureto select for a combination of different plant characteristics that maybe desirable. Such techniques may be used to identify novel quantitativetrait loci (QTLs). By way of example, plants may be selected based ongrowth rate, size (including but not limited to weight, height, leafsize, stem size, branching pattern, or the size of any part of theplant), general health, survival, tolerance to adverse physicalenvironments and/or any other characteristic, as described hereinbefore.

Further non-limiting examples include selecting plants based on: speedof seed germination; quantity of biomass produced; increased root,and/or leaf/shoot growth that leads to an increased yield (pods and/orbeans) or biomass production; effects on plant growth that results in anincreased seed yield for a crop; effects on plant growth which result inan increased pod yield; effects on plant growth that lead to anincreased resistance or tolerance to disease including fungal, viral orbacterial diseases or to pests such as insects, mites or nematodes inwhich damage is measured by decreased foliar symptoms such as theincidence of bacterial or fungal lesions, or area of damaged foliage orreduction in the numbers of nematode cysts or galls on plant roots, orimprovements in plant yield in the presence of such plant pests anddiseases; effects on plant growth that lead to increased metaboliteyields; effects on plant growth that lead to improved aesthetic appealwhich may be particularly important in plants grown for their form,color or taste, for example the color intensity of bean leaves, or thetaste of said leaves.

Molecular Breeding Evaluation Techniques

Selection of plants based on phenotypic or genotypic information may beperformed using techniques such as, but not limited to: high through-putscreening of chemical components of plant origin, sequencing techniquesincluding high through-put sequencing of genetic material, differentialdisplay techniques (including DDRT-PCR, and DD-PCR), nucleic acidmicroarray techniques, RNA-seq (Whole Transcriptome Shotgun Sequencing),qRTPCR (quantitative real time PCR).

In one embodiment, the evaluating step of a plant breeding programinvolves the identification of desirable traits in progeny plants.Progeny plants can be grown in, or exposed to conditions designed toemphasize a particular trait (e.g. drought conditions for droughttolerance, lower temperatures for freezing tolerant traits). Progenyplants with the highest scores for a particular trait may be used forsubsequent breeding steps.

In some embodiments, plants selected from the evaluation step canexhibit a 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 120% or more improvement in aparticular plant trait compared to a control plant.

In other embodiments, the evaluating step of plant breeding comprisesone or more molecular biological tests for genes or other markers. Forexample, the molecular biological test can involve probe hybridizationand/or amplification of nucleic acid (e.g., measuring nucleic aciddensity by Northern or Southern hybridization, PCR) and/or immunologicaldetection (e.g., measuring protein density, such as precipitation andagglutination tests, ELISA (e.g., Lateral Flow test or DAS-ELISA),Western blot, Radioimmune Assay (RIA), immune labeling, immunosorbentelectron microscopy (ISEM), and/or dot blot).

The procedure to perform a nucleic acid hybridization, an amplificationof nucleic acid (e.g., PCT, RT-PCR) or an immunological detection (e.g.,precipitation and agglutination tests, ELISA (e.g., Lateral Flow test orDAS-ELISA), Western blot, RIA, immunogold or immunofluorescent labeling,immunosorbent electron microscopy (ISEM), and/or dot blot tests) areperformed as described elsewhere herein and well-known by one skilled inthe art.

In one embodiment, the evaluating step comprises PCR (semi-quantitativeor quantitative), wherein primers are used to amplify one or morenucleic acid sequences of a desirable gene, or a nucleic acid associatedwith said gene or a desirable trait (e.g., a co-segregating nucleicacid, or other marker).

In another embodiment, the evaluating step comprises immunologicaldetection (e.g., precipitation and agglutination tests, ELISA (e.g.,Lateral Flow test or DAS-ELISA), Western blot, RIA, immuno labeling(gold, fluorescent, or other detectable marker), immunosorbent electronmicroscopy (ISEM), and/or dot blot), wherein one or more gene ormarker-specific antibodies are used to detect one or more desirableproteins. In one embodiment, said specific antibody is selected from thegroup consisting of polyclonal antibodies, monoclonal antibodies,antibody fragments, and combination thereof.

Reverse Transcription Polymerase Chain Reaction (RT-PCR) can be utilizedin the present disclosure to determine expression of a gene to assistduring the selection step of a breeding scheme. It is a variant ofpolymerase chain reaction (PCR), a laboratory technique commonly used inmolecular biology to generate many copies of a DNA sequence, a processtermed “amplification”. In RT-PCR, however, RNA strand is first reversetranscribed into its DNA complement (complementary DNA, or cDNA) usingthe enzyme reverse transcriptase, and the resulting cDNA is amplifiedusing traditional or real-time PCR.

RT-PCR utilizes a pair of primers, which are complementary to a definedsequence on each of the two strands of the cDNA. These primers are thenextended by a DNA polymerase and a copy of the strand is made after eachcycle, leading to logarithmic amplification.

RT-PCR includes three major steps. The first step is the reversetranscription (RT) where RNA is reverse transcribed to cDNA using areverse transcriptase and primers. This step is very important in orderto allow the performance of PCR since DNA polymerase can act only on DNAtemplates. The RT step can be performed either in the same tube with PCR(one-step PCR) or in a separate one (two-step PCR) using a temperaturebetween 40° C. and 60° C., depending on the properties of the reversetranscriptase used.

The next step involves the denaturation of the dsDNA at 95° C., so thatthe two strands separate and the primers can bind again at lowertemperatures and begin a new chain reaction. Then, the temperature isdecreased until it reaches the annealing temperature which can varydepending on the set of primers used, their concentration, the probe andits concentration (if used), and the cations concentration. The mainconsideration, of course, when choosing the optimal annealingtemperature is the melting temperature (Tm) of the primers and probes(if used). The annealing temperature chosen for a PCR depends directlyon length and composition of the primers. This is the result of thedifference of hydrogen bonds between A—T (2 bonds) and G—C (3 bonds). Anannealing temperature about 5 degrees below the lowest Tm of the pair ofprimers is usually used.

The final step of PCR amplification is the DNA extension from theprimers which is done by the thermostable Taq DNA polymerase usually at72° C., which is the optimal temperature for the polymerase to work. Thelength of the incubation at each temperature, the temperaturealterations and the number of cycles are controlled by a programmablethermal cycler. The analysis of the PCR products depends on the type ofPCR applied. If a conventional PCR is used, the PCR product is detectedusing agarose gel electrophoresis and ethidium bromide (or other nucleicacid staining).

Conventional RT-PCR is a time-consuming technique with importantlimitations when compared to real time PCR techniques. This, combinedwith the fact that ethidium bromide has low sensitivity, yields resultsthat are not always reliable. Moreover, there is an increasedcross-contamination risk of the samples since detection of the PCRproduct requires the post-amplification processing of the samples.Furthermore, the specificity of the assay is mainly determined by theprimers, which can give false-positive results. However, the mostimportant issue concerning conventional RT-PCR is the fact that it is asemi or even a low quantitative technique, where the amplicon can bevisualized only after the amplification ends.

Real time RT-PCR provides a method where the amplicons can be visualizedas the amplification progresses using a fluorescent reporter molecule.There are three major kinds of fluorescent reporters used in real timeRT-PCR, general non-specific DNA Binding Dyes such as SYBR Green I,TaqMan Probes and Molecular Beacons (including Scorpions).

The real time PCR thermal cycler has a fluorescence detection threshold,below which it cannot discriminate the difference between amplificationgenerated signal and background noise. On the other hand, thefluorescence increases as the amplification progresses and theinstrument performs data acquisition during the annealing step of eachcycle. The number of amplicons will reach the detection baseline after aspecific cycle, which depends on the initial concentration of the targetDNA sequence. The cycle at which the instrument can discriminate theamplification generated fluorescence from the background noise is calledthe threshold cycle (Ct). The higher is the initial DNA concentration,the lower its Ct will be.

Other forms of nucleic acid detection can include next generationsequencing methods such as DNA SEQ or RNA SEQ using any known sequencingplatform including, but not limited to: Roche 454, Solexa GenomeAnalyzer, AB SOLiD, Illumina GA/HiSeq, Ion PGM, Mi Seq, among others(Liu et al,. 2012 Journal of Biomedicine and Biotechnology Volume 2012ID 251364; Franca et al., 2002 Quarterly Reviews of Biophysics 35 pg169-200; Mardis 2008 Genomics and Human Genetics vol 9 pg 387-402).

In other embodiments, nucleic acids may be detected with other highthroughput hybridization technologies including microarrays, gene chips,LNA probes, nanoStrings, and fluorescence polarization detection amongothers.

In some embodiments, detection of markers can be achieved at an earlystage of plant growth by harvesting a small tissue sample (e.g., branch,or leaf disk). This approach is preferable when working with largepopulations as it allows breeders to weed out undesirable progeny at anearly stage and conserve growth space and resources for progeny whichshow more promise. In some embodiments the detection of markers isautomated, such that the detection and storage of marker data is handledby a machine. Recent advances in robotics have also led to full serviceanalysis tools capable of handling nucleic acid/protein markerextractions, detection, storage and analysis.

Quantitative Trait Loci

Breeding schemes of the present application can include crosses betweendonor and recipient plants. In some embodiments said donor plantscontain a gene or genes of interest which may confer the plant with adesirable phenotype. The recipient line can be an elite line or cultivarhaving certain favorite traits such for commercial production. In oneembodiment, the elite line may contain other genes that also impart saidline with the desired phenotype. When crossed together, the donor andrecipient plant may create a progeny plant with combined desirable lociwhich may provide quantitatively additive effect of a particularcharacteristic. In that case, QTL mapping can be involved to facilitatethe breeding process.

A QTL (quantitative trait locus) mapping can be applied to determine theparts of the donor plant's genome conferring the desirable phenotype,and facilitate the breeding methods. Inheritance of quantitative traitsor polygenic inheritance refers to the inheritance of a phenotypiccharacteristic that varies in degree and can be attributed to theinteractions between two or more genes and their environment. Though notnecessarily genes themselves, quantitative trait loci (QTLs) arestretches of DNA that are closely linked to the genes that underlie thetrait in question. QTLs can be molecularly identified to help mapregions of the genome that contain genes involved in specifying aquantitative trait. This can be an early step in identifying andsequencing these genes.

Typically, QTLs underlie continuous traits (those traits that varycontinuously, e.g. yield, height, level of resistance to virus, etc.) asopposed to discrete traits (traits that have two or several charactervalues, e.g. smooth vs. wrinkled peas used by Mendel in hisexperiments). Moreover, a single phenotypic trait is usually determinedby many genes. Consequently, many QTLs are associated with a singletrait.

A quantitative trait locus (QTL) is a region of DNA that is associatedwith a particular phenotypic trait—these QTLs are often found ondifferent chromosomes. Knowing the number of QTLs that explainsvariation in the phenotypic trait tells about the genetic architectureof a trait. It may tell that a trait is controlled by many genes ofsmall effect, or by a few genes of large effect or by a several genes ofsmall effect and few genes of larger effect.

Another use of QTLs is to identify candidate genes underlying a trait.Once a region of DNA is identified as contributing to a phenotype, itcan be sequenced. The DNA sequence of any genes in this region can thenbe compared to a database of DNA for genes whose function is alreadyknown.

In a recent development, classical QTL analyses are combined with geneexpression profiling i.e., by DNA microarrays. Such expression QTLs(e-QTLs) describes cis- and trans-controlling elements for theexpression of often disease-associated genes. Observed epistatic effectshave been found beneficial to identify the gene responsible by across-validation of genes within the interacting loci with metabolicpathway and scientific literature databases.

QTL mapping is the statistical study of the alleles that occur in alocus and the phenotypes (physical forms or traits) that they produce(see, Meksem and Kahl, The handbook of plant genome mapping: genetic andphysical mapping, 2005, Wiley-VCH, ISBN 3527311165, 9783527311163).Because most traits of interest are governed by more than one gene,defining and studying the entire locus of genes related to a trait giveshope of understanding what effect the genotype of an individual mighthave in the real world.

Statistical analysis is required to demonstrate that different genesinteract with one another and to determine whether they produce asignificant effect on the phenotype. QTLs identify a particular regionof the genome as containing a gene that is associated with the traitbeing assayed or measured. They are shown as intervals across achromosome, where the probability of association is plotted for eachmarker used in the mapping experiment.

To begin, a set of genetic markers must be developed for the species inquestion. A marker is an identifiable region of variable DNA. Biologistsare interested in understanding the genetic basis of phenotypes(physical traits). The aim is to find a marker that is significantlymore likely to co-occur with the trait than expected by chance, that is,a marker that has a statistical association with the trait. Ideally,they would be able to find the specific gene or genes in question, butthis is a long and difficult undertaking. Instead, they can more readilyfind regions of DNA that are very close to the genes in question. When aQTL is found, it is often not the actual gene underlying the phenotypictrait, but rather a region of DNA that is closely linked with the gene.

For organisms whose genomes are known, one might now try to excludegenes in the identified region whose function is known with somecertainty not to be connected with the trait in question. If the genomeis not available, it may be an option to sequence the identified regionand determine the putative functions of genes by their similarity togenes with known function, usually in other genomes. This can be doneusing BLAST, an online tool that allows users to enter a primarysequence and search for similar sequences within the BLAST database ofgenes from various organisms.

Another interest of statistical geneticists using QTL mapping is todetermine the complexity of the genetic architecture underlying aphenotypic trait. For example, they may be interested in knowing whethera phenotype is shaped by many independent loci, or by a few loci, andhow those loci interact. This can provide information on how thephenotype may be evolving.

Molecular markers are used for the visualization of differences innucleic acid sequences. This visualization is possible due to DNA-DNAhybridization techniques (RFLP) and/or due to techniques using thepolymerase chain reaction (e.g. STS, SNPs, microsatellites, AFLP). Alldifferences between two parental genotypes will segregate in a mappingpopulation based on the cross of these parental genotypes. Thesegregation of the different markers may be compared and recombinationfrequencies can be calculated. The recombination frequencies ofmolecular markers on different chromosomes are generally 50%. Betweenmolecular markers located on the same chromosome the recombinationfrequency depends on the distance between the markers. A lowrecombination frequency usually corresponds to a low distance betweenmarkers on a chromosome. Comparing all recombination frequencies willresult in the most logical order of the molecular markers on thechromosomes. This most logical order can be depicted in a linkage map(Paterson, 1996, Genome Mapping in Plants. R. G. Landes, Austin.). Agroup of adjacent or contiguous markers on the linkage map that isassociated to a reduced disease incidence and/or a reduced lesion growthrate pinpoints the position of a QTL.

The nucleic acid sequence of a QTL may be determined by methods known tothe skilled person. For instance, a nucleic acid sequence comprisingsaid QTL or a resistance-conferring part thereof may be isolated from adonor plant by fragmenting the genome of said plant and selecting thosefragments harboring one or more markers indicative of said QTL.Subsequently, or alternatively, the marker sequences (or parts thereof)indicative of said QTL may be used as (PCR) amplification primers, inorder to amplify a nucleic acid sequence comprising said QTL from agenomic nucleic acid sample or a genome fragment obtained from saidplant. The amplified sequence may then be purified in order to obtainthe isolated QTL. The nucleotide sequence of the QTL, and/or of anyadditional markers comprised therein, may then be obtained by standardsequencing methods.

One or more such QTLs associated with a desirable trait in a donor plantcan be transferred to a recipient plant to incorporate the desirabletrait(s) into progeny plants by transferring and/or breeding methods.

In one embodiment, an advanced backcross QTL analysis (AB-QTL) is usedto discover the nucleotide sequence or the QTLs responsible for theresistance of a plant. Such method was proposed by Tanksley and Nelsonin 1996 (Tanksley and Nelson, 1996, Advanced backcross QTL analysis: amethod for simultaneous discovery and transfer of valuable QTL fromun-adapted germplasm into elite breeding lines. Theor Appl Genet92:191-203) as a new breeding method that integrates the process of QTLdiscovery with variety development, by simultaneously identifying andtransferring useful QTL alleles from un-adapted (e.g., land races, wildspecies) to elite germplasm, thus broadening the genetic diversityavailable for breeding. AB-QTL strategy was initially developed andtested in tomato, and has been adapted for use in other crops includingrice, maize, wheat, pepper, barley, and bean. Once favorable QTL allelesare detected, only a few additional marker-assisted generations arerequired to generate near isogenic lines (NILs) or introgression lines(ILs) that can be field tested in order to confirm the QTL effect andsubsequently used for variety development.

Isogenic lines in which favorable QTL alleles have been fixed can begenerated by systematic backcrossing and introgressing of marker-defineddonor segments in the recurrent parent background. These isogenic linesare referred to as near isogenic lines (NILs), introgression lines(ILs), backcross inbred lines (BILs), backcross recombinant inbred lines(BCRIL), recombinant chromosome substitution lines (RCSLs), chromosomesegment substitution lines (CSSLs), and stepped aligned inbredrecombinant strains (STAIRSs). An introgression line in plant molecularbiology is a line of a crop species that contains genetic materialderived from a similar species. ILs represent NILs with relatively largeaverage introgression length, while BILs and BCRILs are backcrosspopulations generally containing multiple donor introgressions per line.As used herein, the term “introgression lines or ILs” refers to plantlines containing a single marker defined homozygous donor segment, andthe term “pre-ILs” refers to lines which still contain multiplehomozygous and/or heterozygous donor segments.

To enhance the rate of progress of introgression breeding, a geneticinfrastructure of exotic libraries can be developed. Such an exoticlibrary comprises a set of introgression lines, each of which has asingle, possibly homozygous, marker-defined chromosomal segment thatoriginates from a donor exotic parent, in an otherwise homogenous elitegenetic background, so that the entire donor genome would be representedin a set of introgression lines. A collection of such introgressionlines is referred as libraries of introgression lines or IL libraries(ILLs). The lines of an ILL cover usually the complete genome of thedonor, or the part of interest. Introgression lines allow the study ofquantitative trait loci, but also the creation of new varieties byintroducing exotic traits. High resolution mapping of QTL using ILLsenable breeders to assess whether the effect on the phenotype is due toa single QTL or to several tightly linked QTL affecting the same trait.In addition, sub-ILs can be developed to discover molecular markerswhich are more tightly linked to the QTL of interest, which can be usedfor marker-assisted breeding (MAB). Multiple introgression lines can bedeveloped when the introgression of a single QTL is not sufficient toresult in a substantial improvement in agriculturally important traits(Gur and Zamir, Unused natural variation can lift yield barriers inplant breeding, 2004, PLoS Biol.; 2(10):e245).

Tissue Culture

As is well known in the art, tissue culture of bean can be used for thein vitro regeneration of a bean plant. Tissue culture of various tissuesof beans and regeneration of plants therefrom is well known and widelypublished. For example, reference may be had to McClean, P., Grafton, K.F., “Regeneration of dry bean (Phaseolus vulgaris) via organogenesis,”Plant Sci., 60, 117-122 (1989); Mergeai, G., Baudoin, J. P.,“Development of an in vitro culture method for heart-shaped embryo inPhaseolus vulgaris,” B.I.C. Invit. Papers 33, 115-116 (1990);Vanderwesthuizen, A. J., Groenewald, E. G., “Root Formation and Attemptsto Establish Morphogenesis in Callus Tissues of Beans (PhaseolusvulgarisL.),” S. Afr. J. Bot. 56, 271-273 (2 April 1990); Benedicic, D.,et al., “The regeneration of Phaseolus vulgaris L. plants from meristemculture,” Abst. 5th I.A.P.T.C. Cong. 1, 91 (#A3-33) (1990); Genga, A.,Allavena, A., “Factors affecting morphogenesis from immature cotyledonsof Phaseolus coccineus L.,” Abst. 5th I.A.P.T.C. Cong. 1, 101 (#A3-75)(1990); Vaquero, F., et al., “Plant regeneration and preliminary studieson transformation of Phaseolus coccineus,” Abst. 5th I.A.P.T.C. Cong. 1,106 (#A3-93) (1990); Franklin, C. I., et al., “Plant Regeneration fromSeedling Explants of Green Bean (Phaseolus-Vulgaris L.) viaOrganogenesis,” Plant Cell Tissue Org. Cult., 24, 199-206 (3 March1991); Malik, K. A., Saxena, P. K., “Regeneration in Phaseolus-vulgarisL.—Promotive Role of N6 Benzylaminopurine in Cultures from JuvenileLeaves,” Planta, 184(1), 148-150 (1991); Genga, A., Allavena, A.,“Factors affecting morphogenesis from immature cotyledons of Phaseoluscoccineus L.,” Plant Cell Tissue Org. Cult., 27, 189-196 (1991); Malik,K. A., Saxena, P. K., “Regeneration in Phaseolus vulgarisL.—High-Frequency Induction of Direct Shoot Formation in IntactSeedlings by N-6-Benzylaminopurine and Thidiazuron,” 186, 384-389 (3Feb. 1992); Malik, K. A., Saxena, P. K., “Somatic Embryogenesis andShoot Regeneration from Intact Seedlings of Phaseolus acutifolius A., P.aureus (L.) Wilczek, P. coccineus L., and P. wrightii L.,” Pl. Cell.Rep., 11, 163 168 (3 Apr. 1992); Chavez, J., et al., “Development of anin vitro culture method for heart shaped embryo in Phaseoluspolyanthus,” B.I.C. Invit. Papers 35, 215-216 (1992); Munoz-Florez, L.C., et al., “Finding out an efficient technique for inducing callus fromPhaseolus microspores,” B.I.C. Invit. Papers 35, 217-218 (1992);Vaquero, F., et al., “A Method for Long-Term Micropropagation ofPhaseolus coccineus L.,” L. Pl. Cell. Rep., 12, 395-398 (7-8 May 1993);Lewis, M. E., Bliss, F. A., “Tumor Formation and beta-GlucuronidaseExpression in Phaseolus vulgaris L. Inoculated with AgrobacteriumTumefaciens,” Journal of the American Society for Horticultural Science,119, 361-366 (2 March 1994); Song, J. Y., et al., “Effect of auxin onexpression of the isopentenyl transferase gene (ipt) in transformed bean(Phaseolus vulgarisL.) single-cell clones induced by Agrobacteriumtumefaciens C58,” J. Plant Physiol. 146, 148-154 (1-2 May 1995). It isclear from the literature that the state of the art is such that thesemethods of obtaining plants are routinely used and have a very high rateof success. Thus, another aspect of this disclosure is to provide cellswhich upon growth and differentiation produce bean plants having thephysiological and morphological characteristics of bean plant WILLS.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as pods, beans, embryos, pollen, flowers, seeds,leaves, stems, roots, root tips, anthers, pistils, meristematic cells,axillary buds, ovaries, seed coat, endosperm, hypocotyls, cotyledons andthe like. Means for preparing and maintaining plant tissue culture arewell known in the art. By way of example, a tissue culture comprisingorgans has been used to produce regenerated plants. U.S. Pat. Nos.5,959,185, 5,973,234, and 5,977,445 describe certain techniques, thedisclosures of which are incorporated herein by reference.

EXAMPLES

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

Example 1 Development of Bean Plant WILLS

Development of bean plant WILLS

Breeding History:

Garden bean cultivar WILLS has superior characteristics and wasdeveloped from an initial cross that was made in Immokalee, Fla., in agreenhouse, in the fall. In the first year of development, the cross wasmade between two proprietary lines under stake numbers BEA3188PL(female) and BEA3433PL (male), the F₁ generation was harvested in Aprilin the greenhouse located in Sun Prairie, Wis., in plot W3411-7, and theF₂ selection was made from the self-pollinated F1 plants in July nearColoma, Wis., in plot H309721. In the second year, the F₃ selection wasmade from the self-pollinated F2 plants in February, near Los Mochis,Mexico, in plot M41868 and the F₄ selection was made from theself-pollinated F3 plants in July near Coloma, Wis., in plot H404827. Inthe third year, the F₅ selection was made from the self-pollinated F4plants in February near Los Mochis, Mexico, in plot M50507 and the F₆selection was made from the self-pollinated F5 plants in July nearColoma, Wis., in plot H503701. In the fourth year, the F₇ generationobtained from the self-pollinated F6 plants was bulked in February nearLos Mochis, Mexico, in plot M62939. In the fifth year, the F₈ generationobtained from the self-pollinated F7 plants was bulked in February nearLos Mochis, Mexico, in plot M72202 and the F₉ generation was harvestedas 100 single plants from the self-pollinated F8 plants in September inTwin Falls, Id., in plot T704291. In the sixth year, the F₁₀ generationobtained from the self-pollinated F9 plants was bulked by progeny row inFebruary, near Los Mochis, Mexico, in plot M83601-648. The line wassubsequently designated WILLS.

Bean plant WILLS is similar to bean plant ‘Wyatt’ (U.S. Pat. No.8,173,877 B2). Wyatt is a commercial bean variety. As shown in Tables 1,2, and 3, while similar to bean plant ‘Wyatt’, there are significantdifferences including bean plant WILLS is resistant to Colletotrichumlindemuthianum (Anthracnose Races 7 and 73), while bean cultivar ‘Wyatt’is susceptible.

Bean plant WILLS is a 55-day maturity bean with uniform medium darkgreen pods on an upright plant structure (habit). The pods are verystraight and smooth and are borne in the upper one-half of the plant.The majority of the pods are in the 3, 4, and 5 sieve range. Bean plantWILLS is a determinate plant and is resistant to Bean common mosaicvirus (BCMV I-gene), Beet curly top virus (BCTV), Colletotrichumlindemuthianum (Anthracnose Races 7 and 73), and Pseudomonas syringae pvsyringae.

The bean plant WILLS has shown uniformity and stability for the traits,within the limits of environmental influence for the traits as describedin the following Variety Descriptive Information. No variant traits havebeen observed or are expected for agronomical important traits in beanplant WILLS.

Bean plant WILLS, has the following morphologic and othercharacteristics, as compared to ‘Wyatt’ (based primarily on datacollected in Sun Prairie, Wis., all experiments done under the directsupervision of the applicants).

TABLE 1 Variety Descriptive Information Variety WILLS WYATT MarketMaturity: Days to edible pods 55 54 Plant: Habit Determinate DeterminatePod position Medium-High Medium-High Bush form High Bush High BushLeaves: Surface: Semi-glossy Semi-glossy Size: Medium Medium Color:Medium Medium dark-green dark-green Anthocyanin Pigment: Flower AbsentAbsent Stem Absent Absent Pods Absent Absent Seeds Absent Absent LeavesAbsent Absent Petioles Absent Absent Peduncles Absent Absent NodesAbsent Absent Flower color: Color of standard White White Color of wingsWhite White Color of keel White White Pods (edible maturity): Exteriorcolor Medium Medium dark-green dark-green Cross section pod shape RoundRound Creaseback Present Present Pubescence Sparse Sparse ConstrictionNone None Fiber Sparse Sparse Number of seeds/pods  7  7 Suture stringAbsent Absent Seed development Medium Medium Machine harvest AdaptedAdapted Distribution of sieve size at optimum maturity 7.34 mm to 8.34mm - Sieve 3 10 30 8.34 mm to 9.53 mm - Sieve 4 70 60 9.53 mm to 10.72mm - Sieve 5 20 10 Seed Color: Seed coat luster Semi-shiny Semi-shinySeed coat Monochrome Monochrome Primary color White Green Seed coatpattern Solid Solid Hilar ring Absent Absent Seed Shape and Size Hilumview Oval Oval Cross section Oval Oval Side view Oval Oval Seed size(g/100 seeds) 26 25 Disease Resistance Bean Common Mosaic VirusResistant Resistant (BCMV I gene) Beet Curly Top Virus (BCTV) ResistantResistant Pseudomonas syringae pv Resistant Resistant syringaeColletotrichum lindemuthianum Resistant Susceptible (Anthracnose Races 7and 73)

Example 2 Field Trials Characteristics of Bean Plant WILLS Field TrialsCharacteristics of Bean Plant WILLS

In Table 2 and 3, the traits and characteristics of bean plant WILLS arecompared to the ‘Wyatt’ variety of beans. The data was collected fromtwo field trial locations at Coloma, Wis. and at Sun Prairie, Wis. Allexperiments were done under the direct supervision of the applicants.

In Tables 2 and 3, the first column shows the “trial location”. Thesecond column shows the “planting date” when seeds were planted in thefield. The third line shows the “harvest date” when the evaluations weredone. The fourth line shows “Trait”, “Variety Name”, and the “CheckVariety Name”. The fifth line shows the “plant height” in inches. Thesixth line shows the “plant width” in inches. The seventh line indicatesthe “plant habit” (structure) with 1=prone (or sprawling) and 9=upright(or erect). The eighth line indicates the “pod length” in millimeters.The ninth line shows the relative “pod color” with 1=light and 9=dark.The tenth line shows the relative “maturity” (the number of days toedible pods). The eleventh line shows the “sieve 1-3%” i.e., thepercentage of pods <8.34 mm in diameter. The twelfth line shows the“sieve 4%” i.e., the percentage of pods between 8.34 mm to 9.53 mm indiameter. The thirteenth column shows the “sieve 5%” i.e., thepercentage of pods >9.53 mm in diameter.

TABLE 2 Variety Characteristics from Field Trials Field Trials TrialLocation Coloma, WI Planting Date May 29 May 29 Harvest date July 26July 26 Trait WILLS Wyatt Plant Height 17 18 Plant Width 18 18 PlantHabit 7 7 Pod Length 145 150 Pod Color 7 8 Maturity 58 58 Sieve 1-3% 168 Sieve 4% 37 67 Sieve 5% 47 25

TABLE 3 Variety Characteristics from Field Trials Field Trials TrialLocation Sun Prairie, WI Planting Date June 7 June 7 Harvest date August7 August 7 Trait WILLS Wyatt Plant Height 19 21 Plant Width 19 21 PlantHabit 7 7 Pod Length 145 165 Pod Color 7 7 Maturity 56 56 Sieve 1-3% 2222 Sieve 4% 30 41 Sieve 5% 48 37

Deposit Information

A deposit of the bean seeds of this disclosure is maintained byHM.CLAUSE, INC., 260 Cousteau Place, Suite 100, Davis, Calif. 95618. Inaddition, a sample of the bean seed of this disclosure has beendeposited with the National Collections of Industrial, Food and MarineBacteria (NCIMB), 23 St Machar Drive, Aberdeen, Scotland, AB24 3RY,United Kingdom.

To satisfy the enablement requirements of 35 U.S.C. 112, and to certifythat the deposit of the isolated strain of the present disclosure meetsthe criteria set forth in 37 C.F.R. 1.801-1.809, Applicants hereby makethe following statements regarding the deposited bean plant WILLS(deposited as NCIMB Accession No. ______,):

-   1. During the pendency of this application, access to the disclosure    will be afforded to the Commissioner upon request;-   2. All restrictions on availability to the public will be    irrevocably removed upon granting of the patent under conditions    specified in 37 CFR 1.808;-   3. The deposit will be maintained in a public repository for a    period of 30 years or 5 years after the last request or for the    effective life of the patent, whichever is longer;-   4. A test of the viability of the biological material at the time of    deposit will be conducted by the public depository under 37 C.F.R.    1.807; and-   5. The deposit will be replaced if it should ever become    unavailable.    Access to this deposit will be available during the pendency of this    application to persons determined by the Commissioner of Patents and    Trademarks to be entitled thereto under 37 C.F.R. § 1.14 and 35    U.S.C. § 122. Upon allowance of any claims in this application, all    restrictions on the availability to the public of the variety will    be irrevocably removed by affording access to a deposit of at least    2,500 seeds of the same variety with the NCIMB.

INCORPORATION BY REFERENCE

All references, articles, publications, patents, patent publications,and patent applications cited herein are incorporated by reference intheir entireties for all purposes.

However, mention of any reference, article, publication, patent, patentpublication, and patent application cited herein is not, and should notbe taken as an acknowledgment or any form of suggestion that theyconstitute valid prior art or form part of the common general knowledgein any country in the world.

What is claimed is:
 1. A seed of bean plant designated WILLS, wherein arepresentative sample of seed of said bean plant having been depositedunder NCIMB No. ______.
 2. A bean plant, a part thereof, or a cellthereof, wherein the bean plant produced by growing the seed of claim 1has all of the physiological and morphological characteristics of beandesignated WILLS deposited under NCIMB No. ______ when grown under thesame environmental conditions.
 3. The bean part of claim 2, wherein thebean part is selected from the group consisting of: a leaf, a flower, apod, an ovule, a seed, a stalk, a root, a rootstock, a scion, an embryo,a stamen, an anther, a pistil, a cell, and pollen.
 4. A tissue cultureof regenerable cells produced from the plant or plant part of claim 2,wherein cells of the tissue culture are produced from a plant part,wherein a plant regenerated from the tissue culture has all of thephysiological and morphological characteristics of bean plant WILLSdeposited under NCIMB No. ______ when grown in the same environmentalconditions.
 5. A bean plant regenerated from the tissue culture of claim4, said plant having all of the physiological and morphologicalcharacteristics of bean plant WILLS, wherein a representative sample ofseed of said bean having been deposited under NCIMB No. ______.
 6. Abean pod or seed produced from the plant of claim
 2. 7. A method forproducing a bean pod, the method comprising a) growing the bean plant ofclaim 2 to produce a bean pod, and b) harvesting said bean pod.
 8. Abean pod produced by the method of claim
 7. 9. A method for producing abean seed, the method comprising (a) crossing a first parent bean plantwith a second parent bean plant and (b) harvesting the resultant beanseed, wherein said first parent bean plant and/or second parent plant isthe bean plant of claim
 2. 10. The bean seed produced by the method ofclaim
 9. 11. A method for producing a bean seed, the method comprising(a) self-pollinating the bean plant of claim 2 and (b) harvesting theresultant bean seed.
 12. A bean seed produced by the method of claim 11.13. A method of vegetatively propagating the bean plant of claim 2, themethod comprising: (a) collecting a part capable of being propagatedfrom the plant of claim 2, (b) regenerating a plant from said part, and(c) harvesting a pod or seed from said regenerated plant.
 14. A plantobtained from the method of claim 13, wherein said plant has all of thephysiological and morphological characteristics of bean plant WILLSdeposited under NCIMB No. ______ when grown under the same environmentalconditions.
 15. A bean pod or seed produced by the method of claim 13.16. A method of producing a bean plant derived from the bean plantWILLS, the method comprising the steps of: (a) crossing the plant ofclaim 2 with itself or a second bean plant to produce a progeny plant;(b) crossing the progeny plant of step (a) with itself or a second beanplant to produce a seed of progeny plant of subsequent generation; (c)growing a progeny plant of the subsequent generation from the seedproduced in step (b); and (d) crossing the progeny plant of step (c)with itself or a second bean plant to produce a bean plant derived fromthe bean plant WILLS.
 17. The method of claim 15 further comprising thestep of: (e) repeating step (c) or (d) for at least one more generationto produce a bean plant derived from the bean plant WILLS.
 18. A methodof producing a plant of bean plant designated WILLS comprising at leastone desired trait, the method comprising: introducing a single locusconversion conferring the desired trait into bean plant designatedWILLS, whereby bean plant designated WILLS comprising the desired traitis produced.
 19. A bean plant, a part thereof, or a cell thereof,produced by the method of claim 18, wherein the plant, the part, or thecell thereof comprises a single locus conversion and essentially all ofthe characteristics of bean plant designated WILLS deposited under NCIMBNo. ______
 20. The plant of claim 19, wherein the single locusconversion confers said plant with a trait selected from the groupconsisting of male sterility, male fertility, herbicide resistance,insect resistance, disease resistance, water stress tolerance, heattolerance, improved shelf life, delayed shelf life and improvednutritional quality.
 21. The plant of claim 19, wherein the single locusconversion is introduced into the plant by the use of recurrentselection, mutation breeding, wherein said mutation breeding selects fora mutation that is spontaneous or artificially induced, backcrossing,pedigree breeding, haploid/double haploid production, marker-assistedselection, genetic transformation, genomic selection, Zinc fingernuclease (ZFN) technology, oligonucleotide directed mutagenesis,cisgenesis, intragenesis, RNA-dependent DNA methylation,agro-infiltration, Transcription Activation-Like Effector Nuclease(TALENs), CRISPR/Cas system, engineered meganuclease, re-engineeredhoming endonuclease, and DNA guided genome editing.
 22. A method ofintroducing a desired trait into bean plant WILLS, the method comprisinga step of: (a) crossing a bean plant WILLS plant grown from bean plantWILLS seed, wherein a representative sample of seed has been depositedunder NCIMB No. ______, with another bean plant that comprises a desiredtrait to produce F1 progeny plants, wherein the desired trait isselected from the group consisting of insect resistance, diseaseresistance, water stress tolerance, heat tolerance, improved shelf lifedelayed shelf life, and improved nutritional quality; (b) selecting oneor more progeny plants that have the desired trait to produce selectedprogeny plants; (c) crossing the selected progeny plants with the beanplant WILLS plants to produce backcross progeny plants; (d) selectingfor backcross progeny plants that have the desired trait and thephysiological and morphological characteristics of bean plant WILLSdeposited under NCIMB No. ______ when grown under the same environmentalcondition to produce selected backcross progeny plants; and (e)repeating steps (c) and (d) three or more times in succession to produceselected fourth or higher backcross progeny plants that comprise thedesired trait and the physiological and morphological characteristics ofbean plant WILLS deposited under NCIMB No. ______ when grown under thesame environmental conditions.
 23. A bean plant produced by the methodof claim 22, wherein the plant has the desired trait and essentially allof the physiological and morphological characteristics of bean plantWILLS deposited under NCIMB No. ______ when grown under the sameenvironmental condition.
 24. A method of producing a bean plant, themethod comprising grafting a rootstock or a scion of the bean plant ofclaim 2 to another bean plant.
 25. A method for producing nucleic acids,the method comprising isolating nucleic acids from the plant of claim 2,or a part, or a cell thereof.